Sequencing data processing method, apparatus, device, medium, and product

Abstract

The application provides a sequencing data processing method, device, equipment, medium and product, and relates to the technical field of nucleic acid sequencing. The method comprises the following steps: obtaining a sequencing read of a target sample, wherein the sequencing read comprises a first marker code and a second marker code; determining whether the information bit coding is correct based on the check bit coding; determining that the information bit coding is incorrect, determining the position of the error base and the corresponding correct base in the information bit coding by using a preset RS algorithm, and generating a corrected first marker code; obtaining the metadata of the target sample from a preset metadata database based on the first marker code; determining whether the second marker code is correct based on the metadata and the corrected first marker code by using a preset hash algorithm; determining that the second marker code is correct, and determining the corrected first marker code and the second marker code as the label of the target sample. The sequencing read comprising two marker codes is double-verified, the sample confusion situation is significantly reduced, and the metadata is included in the marker code, so that the manual recording error is reduced.

Description

Technical Field

[0001] This application relates to the field of nucleic acid sequencing technology, and in particular to a sequencing data processing method, apparatus, equipment, medium and product. Background Technology

[0002] With the rapid development of next-generation sequencing (NGS) technology, the demand for parallel testing of multiple samples in scientific research and clinical practice is becoming increasingly urgent. Traditional single-sample sequencing is costly and inefficient, making it difficult to meet the sequencing needs of large-scale samples. To overcome this bottleneck, multi-sample hybrid sequencing has become the mainstream solution: hundreds or thousands of nucleic acid samples are mixed and fed into the sequencing instrument after library construction, significantly reducing the cost of single-sample sequencing and improving instrument utilization. However, hybrid sequencing also brings a core problem: after the nucleic acid fragments of all samples are randomly fragmented and amplified, the sequencer outputs a massive number of disordered base reads. Without specific identifiers, it is impossible to distinguish which sample a particular read comes from, which can easily lead to sample confusion, failure of data traceability, and even clinical diagnostic errors. Therefore, molecular tagging technology has emerged.

[0003] Existing molecular tagging technologies have developed into several mature schemes, all of which use short nucleic acid sequence tags to differentiate samples. For example, Illumina's index tag technology integrates a 6-10 nt short sequence tag into the 5' or 3' end of the sequencing adapter, with each sample corresponding to a unique index sequence. After mixed sequencing, different samples are distinguished by reading the index tag. Another type is molecular identifier (UMI) technology. For instance, in 10x Genomics single-cell sequencing, a 12 nt UMI tag is added to the cDNA molecule of each single cell for subsequent identification of repetitive sequences introduced by PCR amplification and correction of base errors. Furthermore, some schemes introduce simple verification rules into the tag sequence to reduce the probability of misreading the tag itself during sequencing.

[0004] However, existing molecular tagging technologies have weak error correction capabilities. Most tags rely solely on the sequencer's own quality score to filter errors. If a base misread occurs in the tag sequence (such as A→T and C→G occurring simultaneously), it cannot be identified or corrected, directly leading to sample identification errors. Secondly, the metadata association is loose. The experimental information of the sample (such as batch number and operator ID) needs to be associated through a manually recorded tag-metadata lookup table. This is prone to errors due to manual entry or loss of the table, which can cause the tag and metadata to become disconnected, increasing the risk of sample confusion. Summary of the Invention

[0005] This application provides a sequencing data processing method, apparatus, device, medium, and product to solve the technical problems of weak molecular tag error correction capability and broken association between sample and metadata.

[0006] In a first aspect, this application provides a sequencing data processing method, the method comprising:

[0007] Obtain sequencing reads of the target sample, wherein the sequencing reads include a first marker code and a second marker code, and the first marker code includes: information bit encoding and check bit encoding;

[0008] The correctness of the information bit encoding is determined based on the verification bit encoding.

[0009] In response to the determination that the information bit encoding is incorrect, a preset Reed-Solomon (RS) algorithm is used to determine the position of the incorrect base and the corresponding correct base in the information bit encoding, so as to generate a corrected first marker code;

[0010] Based on the first tag code, the metadata of the target sample is obtained from the preset metadata database. The preset metadata database stores the metadata of multiple samples and the first tag code in association.

[0011] A preset hash algorithm is used to determine whether the second tag code is correct based on the metadata and the corrected first tag code;

[0012] In response to determining that the second tag code is correct, the corrected first tag code and the second tag code are determined as the tags of the target sample, and the tags and the metadata are associated with and stored with the target sample.

[0013] In one possible design, determining whether the information bit encoding is correct based on the check bit encoding includes:

[0014] The check value is calculated based on the information bit encoding using a preset RS algorithm;

[0015] The check value is matched with the check bit code in the first marker code;

[0016] If the match is successful, it is determined that the information bits in the first marker code are correctly encoded;

[0017] If the match fails, it is determined that the information bits in the first marker code are encoded incorrectly.

[0018] In one possible design, the step of using a preset Reed-Solomon (RS) algorithm to determine the position of the erroneous base and the corresponding correct base in the information bit encoding to generate a corrected first marker code includes:

[0019] The incorrect base deviation value is calculated based on the check value and the check bit encoding in the first marker code;

[0020] The incorrect base deviation value is substituted into the preset error position polynomial of the preset RS algorithm to calculate the polynomial result;

[0021] The position of the incorrect base is determined according to the first preset mapping rule and the polynomial result;

[0022] The error base deviation value is substituted into the preset error magnitude polynomial of the preset RS algorithm to calculate the error magnitude.

[0023] The correct base corresponding to the position of the erroneous base is determined according to the second preset mapping rule and the error magnitude.

[0024] Replace the incorrect base with the correct base to generate the corrected first tag code.

[0025] In one possible design, the step of using a preset hash algorithm to determine whether the second tag code is correct based on the metadata and the corrected first tag code includes:

[0026] Generate a mixed number string based on the modified first marker code and the metadata;

[0027] The hash value corresponding to the mixed number string is calculated using a preset hash algorithm;

[0028] Based on the third preset mapping rule, the hash value is converted into the corresponding base to obtain the second tag code corresponding to the corrected first tag code;

[0029] The second marker corresponding to the corrected first marker is matched with the second marker in the sequencing read;

[0030] If a match is successful, the second marker code in the sequencing read is determined to be correct;

[0031] In response to a matching failure, the second marker code in the sequencing read is determined to be incorrect.

[0032] In one possible design, the first marker is located at the 5' end of the sequencing adapter in the sequencing read, and the second marker is located at the 3' end of the insert fragment in the sequencing read.

[0033] In one possible design, the content of guanine (G) and cytosine (C) in the first marker is 40% to 60%;

[0034] And / or, the content of guanine (G) and cytosine (C) in the second marker code is 40%~60%;

[0035] And / or, the second tag code includes degenerate bases.

[0036] In one possible design, prior to acquiring the sequencing data of the target sample, the process further includes:

[0037] In response to receiving a sample tag generation request, an information bit code is randomly obtained from a preset encoding library as the information bit code corresponding to the target sample. The sample tag generation request includes the metadata of the target sample.

[0038] Based on the information bit encoding, a preset RS algorithm is used to generate a check bit encoding, and the information bit encoding and the check bit encoding are merged to determine the first marker code;

[0039] A second tag code is generated based on the first tag code and the metadata using a preset hash algorithm;

[0040] The first and second marker codes are determined as labels for marking the target sample.

[0041] In one possible design, generating the check bit code using a preset RS algorithm based on the information bit code includes:

[0042] The fourth preset mapping rule in the preset RS algorithm is used to convert each base in the information bit encoding into the corresponding mathematical symbol;

[0043] Each of the mathematical symbols is input into a preset polynomial set in the preset RS algorithm to calculate the verification value;

[0044] Based on the fourth preset mapping rule, the check value is converted into the corresponding base to generate the check bit code.

[0045] In one possible design, generating a second tag code based on the first tag code and the metadata using a preset hash algorithm includes:

[0046] Generate a mixed number string based on the first marker code and the metadata;

[0047] The hash value corresponding to the mixed number string is calculated using a preset hash algorithm;

[0048] The hash value is converted into the corresponding base based on the third preset mapping rule to obtain the second tag code.

[0049] In one possible design, after randomly obtaining an information bit code from a preset encoding library as the information bit code corresponding to the target sample, the method further includes:

[0050] The information bit encoding corresponding to the target sample is deleted from the preset encoding library.

[0051] In one possible design, after generating the second token code, the process further includes:

[0052] The first tag code is associated with the metadata and stored in the preset metadata database.

[0053] In one possible design, the metadata of the target sample includes: experimental batch number, timestamp, operator ID, and project number; the method further includes:

[0054] In response to determining that the timestamp has changed, a second token is regenerated based on the first token and metadata including the current timestamp using a preset hash algorithm;

[0055] The first marker code and the regenerated second marker code are determined as labels to mark the target sample.

[0056] Secondly, this application provides a sequencing data processing apparatus, the apparatus comprising:

[0057] The acquisition module is used to acquire sequencing reads of the target sample, wherein the sequencing reads include a first marker code and a second marker code, and the first marker code includes: information bit encoding and check bit encoding;

[0058] The determination module is used to determine whether the information bit encoding is correct based on the check bit encoding;

[0059] The determining module is further configured to, in response to determining that the information bit encoding is incorrect, use a preset Reed-Solomon (RS) algorithm to determine the position of the erroneous base and the corresponding correct base in the information bit encoding, so as to generate a corrected first marker code;

[0060] The acquisition module is further configured to acquire the metadata of the target sample from a preset metadata database based on the first marker code. The preset metadata database stores the metadata of multiple samples and the first marker code in association.

[0061] The determining module is further configured to use a preset hash algorithm to determine whether the second tag code is correct based on the metadata and the corrected first tag code;

[0062] The determining module is further configured to, in response to determining that the second tag code is correct, determine the corrected first tag code and the second tag code as the label of the target sample;

[0063] A storage module is used to associate and store the tags and metadata with the target sample.

[0064] Thirdly, this application provides a sequencing data processing device, the device comprising: a processor, and a memory communicatively connected to the processor;

[0065] The memory is used to store computer-executed instructions;

[0066] The processor is configured to execute computer execution instructions stored in the memory to implement the method as described in any of the first aspects.

[0067] Fourthly, this application provides a computer-readable storage medium storing computer-executable instructions that, when executed by a processor, are used to implement the method as described in any of the first aspects.

[0068] Fifthly, this application provides a computer program product including a computer program that, when executed by a processor, implements the method as described in any of the first aspects.

[0069] The sequencing data processing methods, apparatus, equipment, media, and products provided in this application include tags pre-introduced through molecular biology experiments in the sequencing reads of the target sample, including a first tag code and a second tag code, providing a dual guarantee mechanism for the accuracy and integrity of the data. The first tag code includes bit codes and check bit codes, which not only ensure the correspondence between the tag and the sample but also provide a means of verifying the correctness of the information bit codes, effectively detecting and preventing errors. Using check bit codes to check the correctness of the information bit codes can quickly identify whether errors have occurred during the sequencing process. Detecting errors in the information bit codes at an early stage of the data processing flow can prevent erroneous data from being passed to subsequent steps, reducing the impact of error accumulation on the final result and improving the accuracy and reliability of the entire data processing flow. The RS algorithm is a highly efficient error correction coding algorithm with strong error correction capabilities. When multiple errors occur in the information bit codes, it accurately determines the position of the erroneous bases and recovers the correct base information. By generating a corrected first tag code, not only are errors in the information bit codes corrected, but the integrity and consistency of the original data are also preserved. The corrected first tag code can continue to participate in subsequent data processing, ensuring the smooth operation of the entire process and greatly improving data accuracy and reliability. Since the second tag code is generated based on metadata and the first tag code, when verifying the second tag code, the corresponding metadata is directly retrieved from the preset metadata database. A preset hash algorithm is used in conjunction with the metadata and the corrected first tag code to verify the correctness of the second tag code. This accurately determines whether the second tag code has been tampered with or damaged during data processing, ensuring data integrity and consistency. Furthermore, it allows for a secondary verification of the correctness of the corrected first tag code and metadata. The corrected first and second tag codes are used as labels for the target samples, providing each sample with a unique and clear identifier. Associating and storing the labels and metadata with the target samples achieves comprehensive data association and integration. This facilitates subsequent data querying, analysis, and mining, ensuring data traceability and repeatability. Attached Figure Description

[0070] The accompanying drawings, which are incorporated in and form part of this specification, illustrate embodiments consistent with this application and, together with the description, serve to explain the principles of this application.

[0071] Figure 1 This is an application scenario diagram of the sequencing data processing method provided in one embodiment of this application;

[0072] Figure 2 A flowchart of a sequencing data processing method provided in an embodiment of this application;

[0073] Figure 3 A flowchart illustrating a method for generating sample labels according to an embodiment of this application;

[0074] Figure 4 A flowchart of a sequencing data processing method provided in another embodiment of this application;

[0075] Figure 5 This is a schematic diagram of the structure of a sequencing data processing apparatus provided in one embodiment of this application;

[0076] Figure 6 This is a schematic diagram of the structure of a sequencing data processing device provided in one embodiment of this application.

[0077] The accompanying drawings illustrate specific embodiments of this application, which will be described in more detail below. These drawings and descriptions are not intended to limit the scope of the concept in any way, but rather to illustrate the concept of this application to those skilled in the art through reference to particular embodiments. Detailed Implementation

[0078] Exemplary embodiments will now be described in detail, examples of which are illustrated in the accompanying drawings. When the following description relates to the drawings, unless otherwise indicated, the same numbers in different drawings denote the same or similar elements. The embodiments described in the following exemplary embodiments do not represent all embodiments consistent with this application. Rather, they are merely examples of apparatuses and methods consistent with some aspects of this application as detailed in the appended claims.

[0079] It should be noted that certain software, components, and models may be mentioned in the embodiments of this application. These should be considered as exemplary and are intended only to illustrate the feasibility of implementing the technical solution of this application. However, they do not mean that the applicant has used or necessarily used the solution.

[0080] To clearly understand the technical solution of this application, the solutions of the prior art will be described in detail first.

[0081] The rapid development of next-generation sequencing (NGS) technology allows for the mixing of numerous nucleic acid samples after library construction, significantly reducing the cost per sample and improving instrument utilization. However, after random fragmentation and amplification of nucleic acid fragments, the sequencer outputs a massive number of disordered base reads. Without specific identifiers, it is impossible to distinguish the source of these reads, easily leading to sample confusion, data tracing failure, and even clinical diagnostic errors. This is where molecular tagging technology comes in. Several mature molecular tagging solutions exist, all relying on short nucleic acid sequence labels to differentiate samples. For example, Illumina's index tag technology integrates short sequence tags at the sequencing adapter, using different index sequences to distinguish samples; molecular identifier (UMI) technology adds UMI tags to molecules to identify repetitive sequences and correct base errors. Some solutions also introduce verification rules into the tag sequence to reduce the probability of misreads. However, existing technologies have weak error correction capabilities. Most tags rely on sequencer quality scores to filter errors, and cannot identify and correct base misreads, which can lead to incorrect sample labeling. Secondly, the metadata is loosely linked, and the experimental information of the samples is linked by manually recorded control tables. This is prone to errors in data entry or loss of tables, which can cause the tags to become disconnected from the metadata, increasing the risk of sample confusion.

[0082] Therefore, to address the technical challenges of existing technologies and improve the error correction capability of tags, information bit encoding and check bit encoding are introduced into the first tag code. The information bit encoding serves as the unique index information of the sample, while the check bit encoding is used to verify whether errors exist in the information bit encoding during sequencing. When an error is identified, a pre-defined Reed-Solomon (RS) algorithm is used to determine the position of the erroneous base and its corresponding correct base, which can quickly correct errors in the first tag code, enhancing the error correction capability of molecular tags, reducing the error rate, and correcting PCR repeat and sequencing errors. To avoid the drawbacks of manually recording metadata, the metadata is encoded into the second tag code and stored in a pre-defined metadata database in association with the first tag code. When verifying the second tag code, it is directly retrieved, and a pre-defined hash algorithm is used to verify the correctness of the second tag code in combination with the metadata and the corrected first tag code. This further verifies the association between the sample and the tag, improving the accuracy and reliability of the data. After confirming the correctness of the second tag code, it is used together with the corrected first tag code as the tag for the target sample. The corrected first tag code, the second tag code, and the metadata are bound to the target sample, facilitating subsequent data management and analysis.

[0083] Figure 1 This is an application scenario diagram of the sequencing data processing method provided in one embodiment of this application, such as... Figure 1As shown in the diagram, the application scenario of the sequencing data processing method provided in this embodiment includes: a display screen 101, a sequencing data processing device 102, and a database 103. The sequencing data processing device 102 can be integrated into a drug target prediction device, such as a server or sequencer; this embodiment does not limit this integration.

[0084] Optionally, sequencing data processing can be performed during sequencing or after sequencing is completed; this embodiment does not limit this.

[0085] It should be noted that the display screen 101 can be a display screen on a terminal device, such as a computer screen, and the terminal device is communicatively connected to a server or device that integrates the sequencing data processing device 102.

[0086] Specifically, the user can select which part of the sequencing data to process on the display screen 101, thereby triggering a processing request. After receiving the request, the sequencing data processing device 102 obtains the sequencing read of the target sample. The sequencing read includes a first label and a second label. The first label includes an information bit code and a check bit code. Then, it determines whether the information bit code is correct based on the check bit code. In response to the determination that the information bit code is incorrect, a preset Reed-Solomon (RS) algorithm is used to determine the position of the incorrect base and the corresponding correct base in the information bit code, so as to generate a corrected first label. The sequencing data processing device 102 obtains the metadata of the target sample from a preset metadata database in the database 103 based on the first label. The preset metadata database stores the metadata and first label of multiple samples together. Then, the sequencing data processing device 102 uses a preset hash algorithm based on the metadata and the corrected first label to determine whether the second label is correct. In response to the determination that the second label is correct, the corrected first label and the second label are determined as the label of the target sample, and the label and metadata are placed in the database 103 for associated storage. Users can access and view data in database 103 via display screen 101.

[0087] The technical solution of this application and how the technical solution of this application solves the above-mentioned technical problems are described in detail below with specific embodiments. These specific embodiments can be combined with each other, and the same or similar concepts or processes may not be described again in some embodiments. The embodiments of this application will now be described with reference to the accompanying drawings.

[0088] Figure 2 A flowchart of a sequencing data processing method provided in an embodiment of this application is shown below. Figure 2As shown, the execution entity in this embodiment is a sequencing data processing device. This sequencing data processing device can be implemented through a computer program, or through a medium storing the relevant computer program, such as a USB flash drive and / or optical disc, or it can be integrated into a sequencing data processing device, such as a server or sequencer. The sequencing data processing method provided in this embodiment includes the following steps:

[0089] Step 201: Obtain the sequencing read of the target sample. The sequencing read includes a first marker code and a second marker code. The first marker code includes: information bit encoding and check bit encoding.

[0090] The target sample refers to the biological sample that needs to be sequenced, such as blood, tissue, microorganisms, etc., from which nucleic acid molecules, DNA or RNA are extracted. It is the original research object of the sequencing.

[0091] Among them, a sequencing read refers to a short base sequence output by the sequencer after sequencing the nucleic acid fragments of the target sample. It contains the nucleic acid sequence of the target sample and associated label information.

[0092] The first tag code is a unique molecular tag pre-assigned to the target sample to identify the sample identity. It consists of information bit encoding and check bit encoding.

[0093] Among them, the information bit encoding is a nucleotide sequence (e.g., 8 nucleotides in length) used to carry sample index information. It can generate a massive number of unique identifiers (4 bases) through the permutation and combination of four bases (A / T / C / G). 8 =65536 types), achieving sample differentiation.

[0094] Among them, the check bit code is a nucleotide sequence used for error detection and correction, such as 4 nucleotides in length, which is generated by the RS algorithm based on the information bit code and has a strict mathematical relationship with the information bit code.

[0095] The second marker is a short sequence, such as 8 nucleotides in length, dynamically generated by a hash algorithm based on the first marker and sample metadata, such as batch number and timestamp, and is used for double verification and metadata binding.

[0096] Optionally, if the sequencing data processing device is integrated into the sequencer, it can perform data processing while sequencing the target sample and directly obtain sequencing reads from the sequencer; if the sequencing data processing device is integrated separately into another server or device, a separate database can be set up to store the data output by the sequencer, and sequencing reads can be obtained from it during data processing. This embodiment does not limit this.

[0097] Step 202: Determine whether the information bit encoding is correct based on the check bit encoding.

[0098] Specifically, after obtaining the sequencing read, the complete sequence of the first marker code is extracted from the position of the first marker code in the sequencing read. The information bit code and the check bit code are separated according to the preset rules. The bases of the information bits are converted into numbers, such as A=0, T=1, C=2, G=3. The theoretical check bit code is calculated using the RS algorithm. The theoretical check bit code is compared with the actual check bit code in the sequencing read. If the two are consistent, the information bit code is determined to be correct. If they are inconsistent, the information is determined to be encoded incorrectly.

[0099] Step 203: In response to the determination of an information bit encoding error, the pre-defined Reed-Solomon (RS) algorithm is used to determine the position of the erroneous base and the corresponding correct base in the information bit encoding, so as to generate the corrected first marker code.

[0100] Among them, the preset RS algorithm refers to a predefined error correction coding algorithm that can detect and correct base errors in the information bit coding through the mathematical correlation between the information bit coding and the check bit coding.

[0101] Among them, incorrect bases refer to base misreadings caused by instrument errors, signal interference, etc. during the sequencing process, such as A being misread as T, C being misread as G, or deletion / insertion, which cause the information bit encoding to deviate from the original designed sequence.

[0102] Specifically, when an error is detected in the information bit encoding, a preset RS algorithm is used to first calculate the difference between the actual check bit and the theoretical check bit. This difference is then substituted into a polynomial system within the preset RS algorithm to determine the position of the erroneous base. Next, the difference is substituted into the preset polynomial to calculate the difference between the correct and erroneous bases, thus determining the correct base. The determined correct base is then used to replace the erroneous base, generating the corrected first marker code.

[0103] Understandably, if the information bit encoding is confirmed to be correct, this step can be skipped and the subsequent steps can be performed directly.

[0104] It should be noted that the default RS algorithm can only correct two or fewer erroneous bases. That is, if more than two erroneous base positions are identified, the default RS algorithm cannot be used to correct the first marker code. In this case, the sequencing read can be extracted and manually verified.

[0105] Optionally, after identifying the correct bases, the incorrect bases at the erroneous base positions in the information bit encoding are replaced with the calculated correct bases, such as replacing G with C, to obtain the corrected information bit encoding. Then, the check bit encoding is recalculated based on the corrected information bit encoding. If the calculated check bit encoding matches the check bit encoding in the sequencing read, a corrected first marker code is generated based on the corrected information bit encoding. If the calculated check bit encoding does not match the check bit encoding in the sequencing read, the correction was unsuccessful, and the sequencing read can be extracted for manual verification.

[0106] Step 204: Obtain the metadata of the target sample from the preset metadata database based on the first tag code. The preset metadata database stores the metadata of multiple samples and the first tag code together.

[0107] Among them, the pre-built metadata database refers to a structured database that is pre-built and stored in the sequencing data processing device. Its core function is to establish a one-to-one correspondence between the first marker code and the sample metadata, which is equivalent to a digital identity archive of the sample.

[0108] It is understandable that when generating the label of the target sample, a first tag code is first generated according to a preset method, and then a second tag code is generated based on the first tag code and the metadata of the target sample. The first tag code is then associated with the metadata and stored in a preset database.

[0109] Metadata of the target sample refers to experimental background information related to the target sample, which is structured data describing sample attributes, including but not limited to: experimental batch number, sample preparation timestamp, operator ID, sample source, such as patient ID, tissue type, microbial strain number, project number, sample processing process record, etc.

[0110] Optionally, the associated storage can use an indexing mechanism to bind and store the first identifier of each sample as a unique query key and its corresponding metadata as a value, ensuring that the corresponding metadata can be directly located and retrieved through the first identifier, thus avoiding data corruption. This embodiment does not limit this approach.

[0111] Specifically, after verifying the first tag code, the second verification code is verified. Based on the corrected first tag code, the corresponding metadata is searched in the preset source database and extracted to obtain the metadata of the target sample.

[0112] Step 205: Using a preset hash algorithm based on metadata and the corrected first tag code, determine whether the second tag code is correct.

[0113] Among them, the preset hash algorithm refers to a predefined mathematical algorithm used to map the input metadata and the corrected first tag code into a fixed-length short sequence, such as a custom truncation hash algorithm, a cyclic redundancy check algorithm, and a salted hash algorithm.

[0114] Specifically, after obtaining the metadata, the metadata and the corrected first marker code are input into the formula corresponding to the preset hash algorithm to execute the algorithm. For example, the metadata and the corrected first marker code are first converted into strings that the algorithm can recognize. Then, the preset hash algorithm is used to calculate the hash value of the string. Then, the hash value is converted into a sequence composed of bases through modulo, mapping rules, etc., and compared with the second marker code in the sequencing read. If the comparison is consistent, it means that the second marker code is correct.

[0115] Step 206: In response to determining that the second tag code is correct, the corrected first tag code and second tag code are determined as the tags of the target sample, and the tags and metadata are associated and stored with the target sample.

[0116] Specifically, once the second tag code is confirmed to be correct, the corrected first tag code, such as "ATGCTTACCGAT", and the correct second tag code, such as "CCATCGAC", are extracted and combined according to a preset format to form the final tag of the target sample, for example: ATGCTTACCGAT|CCATCGAC. Then, the tag, metadata, and target sample are bound together and stored as a group, such as by writing them to a database.

[0117] The sequencing data processing method provided in this application includes tags pre-introduced through molecular biology experiments in the sequencing reads of the target sample, including a first tag code and a second tag code, providing a dual guarantee mechanism for the accuracy and integrity of the data. The first tag code includes bit codes and check bit codes, which not only ensure the correspondence between the tag and the sample but also provide a means of verifying the correctness of the information bit codes, effectively detecting and preventing errors. Using check bit codes to check the correctness of the information bit codes can quickly identify whether errors have occurred during the sequencing process. Detecting errors in the information bit codes at an early stage of the data processing flow can prevent erroneous data from being passed to subsequent steps, reducing the impact of error accumulation on the final result and improving the accuracy and reliability of the entire data processing flow. The RS algorithm is a highly efficient error correction coding algorithm with strong error correction capabilities. When multiple errors occur in the information bit codes, it accurately determines the position of the erroneous bases and recovers the correct base information. By generating a corrected first tag code, not only are errors in the information bit codes corrected, but the integrity and consistency of the original data are also preserved. The corrected first tag code can continue to participate in subsequent data processing, ensuring the smooth operation of the entire process and greatly improving data accuracy and reliability. Since the second tag code is generated based on metadata and the first tag code, when verifying the second tag code, the corresponding metadata is directly retrieved from the preset metadata database. A preset hash algorithm is used in conjunction with the metadata and the corrected first tag code to verify the correctness of the second tag code. This accurately determines whether the second tag code has been tampered with or damaged during data processing, ensuring data integrity and consistency. Furthermore, it allows for a secondary verification of the correctness of the corrected first tag code and metadata. The corrected first and second tag codes are used as labels for the target samples, providing each sample with a unique and clear identifier. Associating and storing the labels and metadata with the target samples achieves comprehensive data association and integration. This facilitates subsequent data querying, analysis, and mining, ensuring data traceability and repeatability.

[0118] As an optional implementation, based on the above embodiments, determining whether the information bit encoding is correct based on the check bit encoding includes:

[0119] The check value is calculated based on the information bit encoding using a preset RS algorithm;

[0120] Match the check value with the check bit code in the first marker code;

[0121] If the match is successful, it is determined that the information bits in the first marker code are correctly encoded;

[0122] If a match fails, it is determined that the information bits in the first marker code are encoded incorrectly.

[0123] The check value is a numerical value obtained by calculating the information bit encoding using a preset RS algorithm. It corresponds to the theoretical check value encoding and serves as the theoretical benchmark for judging whether the information bit encoding is correct.

[0124] Specifically, from the sequencing read of the target sample, the storage region of the first marker (e.g., the 5' end of the sequencing adapter) is located, and the complete first marker sequence is extracted according to the preset length of the first marker, such as 12 nucleotides (nt). The first N bases, such as 8 nt, are used as information bits, and the last M bases, such as 4 nt, are used as check bits. The extracted information bit code, such as ATGCTTAC, is converted into a digital sequence according to a fixed rule. For example, the base-to-digit mapping relationship is preset as: A=0, T=1, C=2, G=3, then ATGCTTAC→[0,1,3,2,1,1,0,2]. The digitized information bit code is used as coefficients and substituted into the finite field polynomial in the preset RS algorithm to calculate the check value. After obtaining the check value, it is converted into the corresponding base according to the above fixed rule, thus obtaining the theoretical check bit code. The theoretical check bit code is compared one by one with the check bit code in the sequencing read. If all bases at the same position are the same, the match is successful, indicating that no base misreads, insertions, or deletions occurred during the sequencing process, and the information bit code conforms to the original design sequence, confirming that the information bit code is correct. If one or more bases at the same position are inconsistent, the match fails, indicating that a base misread, insertion, or deletion occurred during the sequencing process, and the information bit code does not conform to the original design sequence, confirming that the information bit code is incorrect.

[0125] For example, the 8nt information bits of the first marker code are encoded as follows: Converting these values ​​to numbers, A=0, T=1, C=2, G=3, these numbers serve as the coefficients of the finite field polynomial: For example, if the encoding is ATCGATCG and the numbers are [0,1,2,3,0,1,2,3], then... The check value is a polynomial value, encoded as the 4nt check bits of the first marker code, set to... , is the result of the calculation of the finite field polynomial at four fixed points in the finite field (e.g., x=1,2,3,4): .

[0126] The sequencing data processing method provided in this application determines the correctness of information bit encoding based on check bit encoding, including: calculating a check value based on the information bit encoding using a preset RS algorithm; matching the check value with the check bit encoding in the first marker code; if the match is successful, the information bit encoding in the first marker code is determined to be correct; if the match fails, the information bit encoding in the first marker code is determined to be incorrect. The RS algorithm is a powerful error correction coding algorithm capable of detecting and correcting a certain number of errors. During sequencing, information bit encoding may be subject to various interferences and produce errors. By calculating the check value using the RS algorithm and matching it with the check bit encoding, errors in the information bit encoding can be detected quickly and accurately, thereby ensuring the integrity and reliability of the data.

[0127] As an optional implementation, based on the above embodiments, a preset Reed-Solomon (RS) algorithm is used to determine the position of the erroneous base and the corresponding correct base in the information bit encoding, so as to generate a corrected first marker code, including:

[0128] The incorrect base deviation value is calculated based on the check value and the check bit encoding in the first marker code;

[0129] Substitute the incorrect base deviation value into the preset error position polynomial of the preset RS algorithm to calculate the polynomial result;

[0130] The position of the incorrect base is determined based on the first preset mapping rule and the polynomial result;

[0131] The error magnitude is calculated by substituting the incorrect base deviation value into the preset error magnitude polynomial of the preset RS algorithm.

[0132] The correct base corresponding to the position of the erroneous base is determined according to the second preset mapping rule and the error magnitude.

[0133] Replace the incorrect base with the correct base to generate the corrected first tag code.

[0134] Among them, the error base deviation value refers to the difference between the value corresponding to the check bit code and the check value. For example, if the check value f(1) = 2 is calculated based on the information code using the preset RS algorithm, but the value corresponding to the check bit code obtained by sequencing is s0 = 3, then the error base deviation value e0 = 3 - 2 = 1.

[0135] The preset error position polynomial refers to the polynomial predefined in the preset RS algorithm used to calculate the position of the erroneous base. It can be a quadratic polynomial, corresponding to a maximum of 2 erroneous bases, and its coefficients are determined by the erroneous base deviation value.

[0136] The first preset mapping rule refers to converting the solution of the polynomial at the error position into the actual base position in the information bit encoding, such as the corresponding rule for the 3rd and 8th positions, such as the root of the polynomial - 1 = the actual position.

[0137] Among them, the preset error magnitude polynomial refers to the set of polynomials used in the preset RS algorithm to calculate the error magnitude, which is solved based on the error base deviation value and the error position. The error magnitude represents the difference between the error base and the correct base.

[0138] The second preset mapping rule refers to the rule for converting error amplitude into correct bases. By combining the sequencing value of the erroneous base, the correct base is deduced. For example, if the error base value + error amplitude = correct base value, it is then mapped to a base.

[0139] Specifically, the calculated check value is extracted, such as a 4nt number sequence. The corresponding base "CGAT" and the check bit in the first marker code encode the 4nt digital sequence obtained from sequencing. = [2,1,3,1], corresponding to the base "CTGT". Calculate the error base deviation values, such as e0 = 2 - 2 = 0, e1 = 1 - 3 = -2 ≡ 3 (modulo 5), e2 = 3 - 0 = 3, e3 = 1 - 1 = 0. Substitute the calculated deviation values ​​into the preset error position polynomial, such as the preset error position polynomial (a quadratic polynomial for 2 error bases): .in, The sum of points in a finite field corresponding to the error location. The product of points in a finite field corresponding to the error location, x i x j Let x be the identifier of the error location in the finite field. Calculate x. i x j , such as x i =2、x j =3. According to the first preset mapping rule, the polynomial calculation result is converted into the position of the erroneous base. For example, if the first preset mapping rule states that the position of the erroneous base = the polynomial calculation result - 1, then error position 1: 2 - 1 = 1, the second bit of the information bit encoding, starting from 0; error position 2: 3 - 1 = 2, the third bit of the information bit encoding, starting from 0. That is, there are errors in the second and third bases of the information bit encoding. Then, the deviation value is substituted into the preset error magnitude polynomial, such as the preset error magnitude polynomial (for 2 erroneous bases): , where a i a j For the error range, The error magnitude is calculated. According to the second mapping rule, the error magnitude is converted into bases. For example, the second preset mapping rule is: Correct base value = Error base sequencing value + Error magnitude, with the value → base mapping: 0 → A, 1 → T, 2 → C, 3 → G, 4 → N. Assume the sequencing value of the error position encoded by the information bit is: 1st bit: rᵢ = 1 (T), 2nd bit: rᵢ = 1 (T), 2nd bit: rᵢ = 1 (T). j =0 (A), calculate the correct base value: 1st bit: 1 + 2 = 3 → corresponding base G, 2nd bit: 0 + 3 = 3 → corresponding base G. Finally, replace the bases at the incorrect base positions with the correct bases to obtain the corrected first tag code.

[0140] Understandably, the entire process described above can be completed automatically by the algorithm, as long as the calculation rules and mapping rules are predefined.

[0141] The sequencing data processing method provided in this application uses a preset Reed-Solomon (RS) algorithm to determine the position of erroneous bases and their corresponding correct bases in the information bit encoding, in order to generate a corrected first marker code. The method includes: calculating an erroneous base deviation value based on a check value and the check bit encoding in the first marker code; substituting the erroneous base deviation value into a preset error position polynomial of the preset RS algorithm to calculate the polynomial result; determining the position of the erroneous base according to a first preset mapping rule and the polynomial result; substituting the erroneous base deviation value into a preset error magnitude polynomial of the preset RS algorithm to calculate the error magnitude; determining the correct base corresponding to the erroneous base position according to a second preset mapping rule and the error magnitude; and replacing the base at the erroneous base position with the correct base to generate the corrected first marker code. By calculating the erroneous base deviation value based on the check value and the check bit encoding, and substituting it into the preset error position polynomial, the position of the erroneous base in the information bit encoding can be accurately determined. Accurate error location is the basis for subsequent effective error correction and can avoid further data damage or error propagation due to inaccurate error location. After identifying the error location, the erroneous base deviation value is substituted into a preset error magnitude polynomial to calculate the error magnitude. Then, the corresponding correct base is determined according to a second preset mapping rule, and finally, the replacement is completed. This series of steps ensures that the erroneous base is accurately corrected to the correct base, restoring the accuracy of the original data to the greatest extent possible, and providing a reliable guarantee for subsequent data analysis and processing.

[0142] As an optional implementation, based on the above embodiments, a preset hash algorithm is used to determine whether the second tag code is correct based on metadata and the corrected first tag code, including:

[0143] Generate a mixed number string based on the corrected first marker code and metadata;

[0144] The hash value corresponding to the mixed number string is calculated using a preset hash algorithm;

[0145] Based on the third preset mapping rule, the hash value is converted into the corresponding base to obtain the second tag code corresponding to the corrected first tag code;

[0146] Match the second marker code corresponding to the corrected first marker code with the second marker code in the sequencing read;

[0147] If a match is successful, it is determined that the second marker code in the sequencing read is correct;

[0148] If a match fails, the second marker code in the sequencing read is determined to be incorrect.

[0149] Among them, the hybrid digital string refers to the continuous digital sequence formed by concatenating the corrected first marker code and metadata into digital form according to preset rules. This sequence serves as the input data for the hash algorithm, ensuring that different types of information (nucleic acid sequences and text metadata) can be processed uniformly by the algorithm.

[0150] A hash value is a fixed-length numerical value obtained by calculating a mixed string of numbers using a preset hash algorithm, such as a 128-bit or 160-bit binary number. It is unique: different inputs correspond to different hash values, and it is unidirectional; the original input cannot be deduced from the hash value.

[0151] The third preset mapping rule refers to the rule for converting hash values ​​into a base sequence of a specific length, i.e., the second marker code, such as one base corresponding to every four binary bits.

[0152] Specifically, the corrected first marker code and metadata are each converted into numeric characters. For example, if the corrected first marker code sequence is ATGCTTACCGAT, it is converted into a numeric string according to the preset base-number mapping rule (A=0, T=1, C=2, G=3): "ATGCTTACCGAT"→0 1 3 2 1 1 0 2 2 3 0 1. The metadata of the target sample is retrieved from the preset metadata database, such as {"batch_id":"B005","timestamp":"2024-06-15","operator_id":"OP-012"}). Text metadata: non-numeric characters are removed, and numbers are retained, such as "B005"→"005", "OP-012"→"012"; timestamp: converted into a numeric string, such as "2024-06-15"→"20240615"; the metadata numeric string is concatenated: "00520240615012". In a fixed order, such as the first tag code string first and the metadata string last, they are concatenated and connected by a separator: 013211022301-00520240615012.

[0153] The mixed number string is input into a preset hash algorithm suitable for short sequence generation, and a hash value is obtained by performing hash calculation, such as: 10110011001011011000111011001010. Based on the third preset mapping rule, the hash value is converted into the corresponding base. For example, the third preset mapping rule is to group the 32-bit binary hash value into 8 groups of 4 bits each, with each group corresponding to 1 base: 0000→A, 0001→T, 0010→C, 0011→G, 0100→A, 0101→T, 0110→C, 0111→G (the remaining combinations are cyclically mapped). The hash value fragment is: 1011 0011 0010 1101 1000 1110 1100 1010. Each group of transformations is 1011→G, 0011→G, 0010→C, 1101→G, 1000→A, 1110→C, 1100→T, 1010→T. Concatenating these fragments yields the second tag code corresponding to the 8nt corrected first tag code: GGCGACTT.

[0154] The second marker corresponding to the corrected first marker is matched base-by-base with the second marker in the sequencing read. If they match perfectly, the match is successful, indicating that the corrected first marker is accurate; the metadata and sample association records are correct; and no substantial errors occurred in the second marker during sequencing, confirming that the second marker in the sequencing read is correct. If there are base mismatches, the match fails, indicating an error in the second marker in the sequencing read. This can trigger an alert mechanism, prompting the operator to check the first marker correction process, the metadata database, or the sample processing procedure, and to re-sequencing if necessary.

[0155] The sequencing data processing method provided in this application uses a preset hash algorithm to determine the correctness of a second marker code based on metadata and a corrected first marker code. The method includes: generating a mixed number string based on the corrected first marker code and metadata; calculating the hash value corresponding to the mixed number string using a preset hash algorithm; converting the hash value into corresponding bases based on a third preset mapping rule to obtain the second marker code corresponding to the corrected first marker code; matching the second marker code corresponding to the corrected first marker code with the second marker code in the sequencing read; determining that the second marker code in the sequencing read is correct if the match is successful; and determining that the second marker code in the sequencing read is incorrect if the match fails. Generating a mixed number string from the corrected first marker code and metadata before calculating the hash value allows for simultaneous consideration of marker code information and related metadata. Metadata may contain important information such as sequencing time, sample source, and sequencing instrument parameters. Combining this information allows for more comprehensive and accurate verification of the correctness of the second marker code, avoiding potential misjudgments that may occur when relying on only a single information source. The hash algorithm is unique and deterministic; the same input will inevitably produce the same output. By precisely calculating the hash value of the mixed digital string and converting it into bases based on a preset mapping rule to obtain the corresponding second marker code, a strict match is then performed with the second marker code in the sequencing read. This precise matching mechanism greatly reduces the probability of misjudgment due to random errors or accidental factors, ensuring a high degree of accuracy in data verification results.

[0156] As an optional implementation, the first marker is located at the 5' end of the sequencing adapter in the sequencing read, and the second marker is located at the 3' end of the insert fragment in the sequencing read.

[0157] Sequencing adapters are short sequences artificially added to both ends of nucleic acid fragments during tag construction. Their function is to enable the nucleic acid fragments to bind to the flow cell of the sequencer and provide sequencing start sites. Their 5' ends are common embedding sites for tag codes.

[0158] The 5' end is the start end of the nucleic acid sequence. In chemical terms, one end of the nucleic acid chain is a 5' phosphate group and the other end is a 3' hydroxyl group. The 5' end is the head of the chain, and the sequence at the 5' end is usually read first during sequencing.

[0159] Among them, the inserted fragment refers to the core nucleic acid fragment that needs to be sequenced extracted from the target sample, such as genomic DNA fragment or cDNA fragment, whose 3' end, that is, the end of the sequence, can be embedded with auxiliary marker codes.

[0160] The 3' end is the end of the nucleic acid sequence. In paired-end sequencing, the sequencer reads from the 5' end to the 3' end once, and then reads from the 3' end to the 5' end once in reverse order, achieving bidirectional coverage of the inserted fragment.

[0161] Specifically, tags can be introduced at designated locations through molecular biology experiments. For example, a sequencing adapter with a first tag code can be synthesized: the adapter sequence consists of a universal adapter backbone plus a first tag code, where the first tag code is located at the 5' end of the adapter. This adapter is then ligated to the 5' end of the insert fragment using DNA ligase, naturally placing the first tag code at the very front of the entire nucleic acid molecule. A second tag code is added to the 3' end of the PCR primers used to amplify the insert fragment. Using the target nucleic acid fragment as a template, PCR amplification is performed using these primers, allowing the second tag code to be integrated into the 3' end of the insert fragment, ultimately forming a structure of insert fragment + 3' end second tag code.

[0162] Understandably, during sequencing, the sequencer first starts sequencing from the 5' end of the nucleic acid fragment. The first region read is the 5' end of the sequencing adapter, so the first marker is preferentially identified and recorded. After completing the 5' end reading, the sequencer reads the end of the insert fragment from the 3' end using either indexed reading or reverse reading of paired-end sequencing.

[0163] The sequencing data processing method provided in this application has a first marker located at the 5' end of the sequencing adapter in the sequencing read, and a second marker located at the 3' end of the insert fragment in the sequencing read. The first and second markers are located at different positions, allowing their generated signals to be detected and identified relatively independently in time and space. This avoids interference between the two marker signals, improves the accuracy and sensitivity of signal detection, and thus enables more accurate reading of sequencing data.

[0164] As an optional implementation, based on the above embodiments, the content of guanine (G) and cytosine (C) in the first marker is 40%~60%;

[0165] And / or, the content of guanine (G) and cytosine (C) in the second marker code is 40%~60%;

[0166] And / or, the second marker code includes degenerate bases.

[0167] Degenerate bases refer to a type of nucleic acid sequence design where a position is not fixed to a single base (A / T / C / G), but contains a mixture of two or more bases. For example, N represents any one of A / T / C / G, Y represents C / T, R represents A / G, etc. This can increase the diversity of marker codes or adapt to the sequence compatibility of different samples.

[0168] It is understandable that the GC content of the first and second marker codes, as well as the inclusion of degenerate bases in the second marker code, can be controlled through molecular biology experiments.

[0169] The sequencing data processing method provided in this application has a first marker code containing 40%–60% guanine (G) and cytosine (C); and / or, a second marker code containing 40%–60% guanine (G) and cytosine (C); and / or, the second marker code includes degenerate bases. Excessively high or low G and C contents can easily lead to the formation of complex secondary structures in the marker code itself, such as hairpin structures and stem-loop structures. These secondary structures can interfere with the binding of the marker code to sequencing primers, enzymes, and other reagents, affecting the sequencing reaction. Appropriate G and C contents can reduce the likelihood of secondary structure formation and ensure the smooth progress of the sequencing reaction. The presence of degenerate bases allows the second marker code to capture more sequence variation information. During sequencing, even if some unknown base variations exist in the sample, the marker code containing degenerate bases can still bind to them, thereby recording this variation information. This helps in discovering rare variations or new genotypes, improving the sensitivity and coverage of sequencing.

[0170] As an optional implementation, based on the above embodiments, before obtaining the sequencing data of the target sample, the following steps are also included:

[0171] In response to receiving a sample label generation request, an information bit code is randomly obtained from a preset encoding library as the information bit code corresponding to the target sample. The sample label generation request includes the metadata of the target sample.

[0172] Based on the information bit encoding, a check bit encoding is generated using a preset RS algorithm, and the information bit encoding and the check bit encoding are merged to determine the first marker code;

[0173] A second tag code is generated based on the first tag code and metadata using a preset hash algorithm;

[0174] The first and second marker codes are used as labels to mark the target samples.

[0175] Among them, the sample label generation request refers to the instruction initiated by the user to create a unique molecular label for the target sample, which includes the target sample's metadata, such as batch number, operator ID, etc., and triggers the label generation process.

[0176] Among them, the pre-built encoding library refers to a pre-built database containing a large number of unique information bit codes, such as 1 million 8nt sequences, which are used to quickly assign sample identifiers.

[0177] Specifically, upon receiving a sample label generation request, the metadata of the target sample is parsed, and its integrity is verified, such as whether any key fields are missing. A preset encoding library is accessed, and one unused information bit code is randomly selected. The assigned information bit code is input into a preset RS algorithm to generate a check code, which is then concatenated in the order of information bit code + check bit code to obtain the complete first label code. The first label code and metadata are input into a preset hash algorithm to generate a second label code. For example, the information bit code and metadata are converted into a string of numbers according to preset rules, then input into the preset hash algorithm to calculate the hash value, and finally converted into the corresponding bases to obtain the second label code. The first and second label codes are combined in a format such as ATGCTTACCGAT:GGC GACTT as the unique molecular tag for the target sample, used to label the target sample.

[0178] Optionally, after obtaining the information and encoding it, update the database fields and record the allocation time and associated sample ID to avoid duplicate allocation.

[0179] Understandably, after generating the tag for the target sample, Xu conducts molecular biology experiments to link the tag to the target sample so that the tag can be read during sequencing.

[0180] The sequencing data processing method provided in this application embodiment further includes, before acquiring the sequencing data of the target sample: in response to receiving a sample tag generation request, randomly obtaining an information bit code from a preset coding library as the information bit code corresponding to the target sample, wherein the sample tag generation request includes the metadata of the target sample; generating a check bit code using a preset RS algorithm based on the information bit code, and merging the information bit code and the check bit code to determine a first tag code; generating a second tag code based on the first tag code and the metadata using a preset hash algorithm; and determining the first tag code and the second tag code as a tag for tagging the target sample. Randomly obtaining the information bit code from the preset coding library as the code corresponding to the target sample ensures that each sample has a unique information bit identifier. The first tag code formed by combining the subsequently generated check bit code further strengthens uniqueness, accurately distinguishes different samples, and avoids sample confusion. The second tag code is generated using a preset hash algorithm based on the first tag code and sample metadata, wherein the metadata includes key information such as sample source and collection time. The second tag code combines the unique code of the sample with important attribute information, identifying the sample from multiple dimensions, making the sample closely associated with the metadata for easy subsequent management. Generate accurate sample labels before acquiring sequencing data to control quality at the source of data generation. Accurate labeling ensures that samples are correctly identified and processed in subsequent stages, preventing the propagation and accumulation of data errors due to incorrect sample labeling.

[0181] As an optional implementation, based on the above embodiments, a check bit code is generated using a preset RS algorithm according to the information bit code, including:

[0182] The fourth preset mapping rule in the preset RS algorithm is used to convert each base in the information bit encoding into the corresponding mathematical symbol;

[0183] Input each mathematical symbol into the preset polynomial set in the preset RS algorithm to calculate the verification value;

[0184] Based on the fourth preset mapping rule, the check value is converted into the corresponding base to generate the check bit code.

[0185] The fourth mapping rule refers to the rule for converting nucleic acid bases (A / T / C / G) to mathematical symbols, such as A→0, T→1, C→2, G→3, so that the base sequence can be processed by the mathematical polynomial system of the preset RS algorithm.

[0186] Among them, the preset polynomial set refers to a set of polynomials used in the preset RS algorithm to calculate the check value. The polynomials take the mathematical symbols after the information bit encoding conversion as input and output the check value corresponding to the check bit length.

[0187] Specifically, according to the fourth mapping rule, the bases in the information encoding are converted into mathematical symbols. For example, the fourth preset mapping rule is A→0, T→1, C→2, G→3. Assuming the information bit encoding is 8nt ATGCTTAC, each base is read and converted sequentially: A→0, T→1, G→3, C→2, T→1, T→1, A→0, C→2, resulting in the corresponding mathematical symbol sequence [0,1, 3, 2, 1, 1, 0, 2]. Each mathematical symbol is input into a preset polynomial set and calculated to obtain a check value, such as a four-bit check value. = [2,3,0,1]. Based on the fourth preset mapping rule, the check value is converted into the corresponding base, such as... = [2,3,0,1]=[CGAT], thus obtaining the check digit code.

[0188] The sequencing data processing method provided in this application generates check bit codes based on information bit encoding using a preset RS algorithm. The method includes: converting each base in the information bit encoding into a corresponding mathematical symbol using a fourth preset mapping rule in the preset RS algorithm; inputting each mathematical symbol into a preset polynomial set in the preset RS algorithm to calculate a check value; and converting the check value back into corresponding bases based on the fourth preset mapping rule to generate a check bit code. After converting the bases in the information bit encoding into mathematical symbols, calculations are performed using a preset polynomial set. Mathematical operations have high precision, enabling accurate and rapid calculation of the check value. The check value is then converted back into bases according to the preset rules to obtain the check bit code. This not only improves the speed and accuracy of tag generation but also effectively prevents data tampering due to the strong correlation of mathematical characteristics.

[0189] As an optional implementation, based on the above embodiments, a preset hash algorithm is used to generate a second tag code based on the first tag code and metadata, including:

[0190] Generate a mixed number string based on the first marker code and metadata;

[0191] The hash value corresponding to the mixed number string is calculated using a preset hash algorithm;

[0192] The hash value is converted into the corresponding base based on the third preset mapping rule to obtain the second tag code.

[0193] Specifically, the first marker code sequence, such as ATGCTTACCGAT, is extracted and converted into a continuous numeric string (013211022301) according to a preset base-number mapping rule (A=0, T=1, C=2, G=3). The extracted metadata, such as {"sample_id":"S001","batch_id":"B005","timestamp":"20240615","operator":"OP012"}, is converted into a metadata numeric string (00100520240615012) according to the rules. These are then concatenated into a mixed numeric string (013211022301#00100520240615012). This mixed numeric string is then input into a preset hash algorithm to calculate the hash value. Finally, according to the third mapping rule, the hash value is converted into the corresponding bases to obtain the second marker code.

[0194] Understandably, this process is similar to the aforementioned process of determining whether the second marker code is correct.

[0195] The sequencing data processing method provided in this application uses a preset hash algorithm to generate a second marker code based on a first marker code and metadata. The method includes: generating a mixed number string based on the first marker code and metadata; calculating the hash value corresponding to the mixed number string using the preset hash algorithm; and converting the hash value into corresponding bases based on a third preset mapping rule to obtain the second marker code. The first marker code itself already possesses a preliminary unique identification function for the sample, while the metadata contains key information such as the sample's source, collection time, and sample type. Integrating this important information allows the mixed number string to cover multiple key features of the sample, greatly increasing its uniqueness. The preset hash algorithm has the characteristic of mapping inputs of arbitrary length to fixed-length outputs, and it is almost impossible to obtain the same output hash value for different inputs. Therefore, the hash value calculated based on the unique mixed number string is also unique. The second marker code obtained after converting the hash value into bases using the third preset mapping rule can provide a unique identifier for each sample, effectively avoiding duplication and confusion of sample identifiers.

[0196] As an optional implementation, based on the above embodiments, after randomly obtaining an information bit code from a preset encoding library as the information bit code corresponding to the target sample, the method further includes:

[0197] Remove the information bit encoding corresponding to the target sample from the preset encoding library.

[0198] Specifically, after randomly obtaining an information bit encoding from the preset encoding library, the encoding is deleted from the preset encoding library to avoid the subsequent reuse of the encoding.

[0199] The sequencing data processing method provided in this application, after randomly obtaining an information bit code from a preset coding library as the information bit code corresponding to the target sample, further includes: deleting the information bit code corresponding to the target sample from the preset coding library. Deleting the obtained code ensures that the information bit code corresponding to each sample is different and unique.

[0200] As an optional implementation, after generating the second marker code based on the above embodiments, the method further includes:

[0201] The first tag code is associated with metadata and stored in a preset metadata database.

[0202] Specifically, the first tag code and metadata are packaged, and an association and index are established and stored in a preset metadata database so that the corresponding metadata can be retrieved using the first tag code in subsequent metadata retrieval. There is a unique association between the first tag code and the metadata.

[0203] The sequencing data processing method provided in this application, after generating the second marker, further includes: associating the first marker with metadata and storing it in a preset metadata database. Associating the first marker with metadata in the preset metadata database centralizes scattered sample-related information, avoiding information dispersion and loss, and facilitating subsequent verification. There is a close logical connection between the first marker and metadata; associative storage ensures their consistency. When the first marker changes, the metadata database can update the associated metadata promptly; and vice versa. For example, if the first marker of a sample needs to be modified for some reason, the associative storage mechanism ensures that the sample-related information recorded in the metadata can also be adjusted accordingly, avoiding data inconsistencies.

[0204] As an optional implementation, based on the above embodiments, the metadata of the target sample includes: experimental batch number, timestamp, operator ID, and project number. The method further includes:

[0205] In response to the determination that the timestamp has changed, a second token is regenerated based on the first token and metadata including the current timestamp using a preset hash algorithm;

[0206] The first marker code and the regenerated second marker code are used as labels to label the target sample.

[0207] It is understandable that the timestamps recorded in the metadata of the target sample, such as the sample preparation time and modification time, may be inconsistent with the initial records. This is because the time may be updated due to adjustments in the experimental procedure or corrections of data entry errors.

[0208] The current timestamp refers to the latest timestamp that takes effect after modification, replacing the old timestamp in the metadata, and serving as the input parameter for regenerating the second tag code.

[0209] Specifically, a monitoring mechanism detects updates to the timestamp in the metadata, automatically triggering the regeneration process of the second tag code. When an operator modifies the timestamp in the sample metadata, such as changing the old timestamp "2024-06-15 09:30:00" to the new timestamp "2024-06-16 10:20:00", the values ​​before and after the change are recorded. The old and new timestamps are compared; if they are inconsistent, it is determined that the timestamp has changed, and a regeneration instruction for the second tag code is generated. The preset metadata database is queried based on the sample ID to obtain the first tag code corresponding to the sample. Following the aforementioned method, the second tag code is regenerated based on the first tag code and the metadata including the current timestamp. The first tag code and the regenerated second tag code are used as labels to mark the target sample.

[0210] It is understandable that for the same sample, if the timestamp changes, the first tag code remains the same, but the second tag code is different.

[0211] The sequencing data processing method provided in this application includes the following metadata for the target sample: experimental batch number, timestamp, operator ID, and project number. The method further includes: in response to a determined change in the timestamp, regenerating a second marker code based on a first marker code and the metadata including the current timestamp using a preset hash algorithm; and determining the first marker code and the regenerated second marker code as tags to label the target sample. Since the second marker code is generated based on the first marker code and the metadata containing the current timestamp, a change in the timestamp will cause a change in the second marker code. This dynamic generation mechanism makes the marker code difficult to predict and forge, further enhancing the data's anti-tampering capability.

[0212] Figure 3 A flowchart of a method for generating sample labels provided in an embodiment of this application is shown below. Figure 3 As shown, the method for generating sample labels provided in this embodiment includes the following steps:

[0213] Step 301: In response to receiving a sample tag generation request, randomly select an information bit code from a preset encoding library as the information bit code corresponding to the target sample. The sample tag generation request includes the metadata of the target sample.

[0214] Step 302: Delete the information bit encoding corresponding to the target sample from the preset encoding library.

[0215] Step 303: Using the fourth preset mapping rule in the preset RS algorithm, each base in the information bit encoding is converted into its corresponding mathematical symbol.

[0216] Step 304: Input each mathematical symbol into the preset polynomial set in the preset RS algorithm to calculate the verification value.

[0217] Step 305: Based on the fourth preset mapping rule, the check value is converted into the corresponding base to generate the check bit code, and the information bit code and the check bit code are merged to determine the first marker code.

[0218] Step 306: Generate a mixed number string based on the first marker code and metadata.

[0219] Step 307: Calculate the hash value corresponding to the mixed number string using a preset hash algorithm.

[0220] Step 308: Based on the third preset mapping rule, the hash value is converted into the corresponding base to obtain the second tag code.

[0221] Step 309: Associate the first tag code with metadata and store it in the preset metadata database.

[0222] Understandably, the above steps are the process of generating a unique label for each sample. After determining the label for each sample, a molecular label is prepared using biological experiments and linked to the sample to obtain a sample containing a first label code and a second label code.

[0223] In this embodiment, the implementation method and technical effect of steps 301-309 are similar to those of the corresponding solutions in the above embodiments, and will not be repeated here.

[0224] Figure 4 A flowchart of a sequencing data processing method provided in another embodiment of this application is shown below. Figure 4 As shown, the sequencing data processing method provided in this embodiment includes specific steps for verifying the first and second marker codes. Therefore, the sequencing data processing method provided in this embodiment includes the following steps:

[0225] Step 401: Obtain the sequencing read of the target sample. The sequencing read includes a first marker code and a second marker code. The first marker code includes: information bit encoding and check bit encoding.

[0226] Step 402: Calculate the check value based on the information bit encoding using the preset RS algorithm.

[0227] Step 403: Match the check value with the check bit code in the first marker code.

[0228] Step 404: If the match fails, it is determined that the information bit encoding in the first marker code is incorrect.

[0229] Step 405: Calculate the incorrect base deviation value based on the check value and the check bit encoding in the first marker code.

[0230] Step 406: Substitute the incorrect base deviation value into the preset error position polynomial of the preset RS algorithm to calculate the polynomial result.

[0231] Step 407: Determine the position of the incorrect base according to the first preset mapping rule and the polynomial result.

[0232] Step 408: Substitute the incorrect base deviation value into the preset error magnitude polynomial of the preset RS algorithm to calculate the error magnitude.

[0233] Step 409: Determine the correct base corresponding to the erroneous base position according to the second preset mapping rule and the error magnitude.

[0234] Step 410: Replace the bases at the incorrect base positions with the correct bases to generate the corrected first tag code.

[0235] Step 411: Obtain the metadata of the target sample from the preset metadata database based on the first tag code. The preset metadata database stores the metadata of multiple samples and the first tag code together.

[0236] Step 412: Generate a mixed number string based on the corrected first marker code and metadata.

[0237] Step 413: Calculate the hash value corresponding to the mixed number string using a preset hash algorithm.

[0238] Step 414: Based on the third preset mapping rule, convert the hash value into the corresponding base to obtain the second tag code corresponding to the corrected first tag code.

[0239] Step 415: Match the second marker code corresponding to the corrected first marker code with the second marker code in the sequencing read.

[0240] Step 416: In response to a successful match, the second marker code in the sequencing read is confirmed to be correct.

[0241] Step 417: In response to determining that the second tag code is correct, the corrected first tag code and second tag code are determined as the tags of the target sample, and the tags and metadata are associated and stored with the target sample.

[0242] In this embodiment, the implementation method and technical effect of steps 401-417 are similar to those of the corresponding solutions in the above embodiments, and will not be repeated here.

[0243] Figure 5 This is a schematic diagram of the structure of a sequencing data processing device provided in an embodiment of this application, as shown below. Figure 5 As shown, the sequencing data processing device provided in this embodiment is located within the sequencing data processing equipment. The sequencing data processing device 50 provided in this embodiment includes: an acquisition module 51, a determination module 52, and a storage module 53.

[0244] The system includes: an acquisition module 51 for acquiring sequencing reads of the target sample, the sequencing reads including a first label and a second label, the first label including an information bit code and a check bit code; a determination module 52 for determining whether the information bit code is correct based on the check bit code; the determination module 52 is also used to determine the position of the erroneous base and the corresponding correct base in the information bit code and generate a corrected first label in response to the determination that the information bit code is incorrect, using a preset Reed-Solomon (RS) algorithm; the acquisition module 51 is also used to acquire metadata of the target sample from a preset metadata database based on the first label, the preset metadata database storing metadata and the first label of multiple samples together; the determination module 52 is also used to determine whether the second label is correct based on the metadata and the corrected first label using a preset hash algorithm; the determination module 52 is also used to determine the corrected first label and the second label as the label of the target sample in response to the determination that the second label is correct; and a storage module 53 for storing the label and metadata in association with the target sample.

[0245] The sequencing data processing device provided in this embodiment can perform... Figure 2 The methods provided in the embodiments are similar in their specific implementation principles and technical effects, and will not be described in detail here.

[0246] Optionally, the determining module 52, when determining whether the information bit encoding is correct based on the check bit encoding, is specifically used to: calculate the check value based on the information bit encoding using a preset RS algorithm; match the check value with the check bit encoding in the first tag code; if the match is successful, determine that the information bit encoding in the first tag code is correct; if the match fails, determine that the information bit encoding in the first tag code is incorrect.

[0247] Optionally, the determining module 52, when using a preset Reed-Solomon (RS) algorithm to determine the position of the erroneous base and the corresponding correct base in the information bit encoding to generate the corrected first tag code, specifically performs the following: calculating the erroneous base deviation value based on the check value and the check bit encoding in the first tag code; substituting the erroneous base deviation value into the preset error position polynomial of the preset RS algorithm to calculate the polynomial result; determining the position of the erroneous base according to the first preset mapping rule and the polynomial result; substituting the erroneous base deviation value into the preset error magnitude polynomial of the preset RS algorithm to calculate the error magnitude; determining the correct base corresponding to the erroneous base position according to the second preset mapping rule and the error magnitude; and replacing the base at the erroneous base position with the correct base to generate the corrected first tag code.

[0248] Optionally, the determining module 52, when determining whether the second marker is correct based on metadata and the corrected first marker using a preset hash algorithm, is specifically configured to: generate a mixed number string based on the corrected first marker and metadata; calculate the hash value corresponding to the mixed number string using a preset hash algorithm; convert the hash value into the corresponding base based on a third preset mapping rule to obtain the second marker corresponding to the corrected first marker; match the second marker corresponding to the corrected first marker with the second marker in the sequencing read; if the match is successful, determine that the second marker in the sequencing read is correct; if the match fails, determine that the second marker in the sequencing read is incorrect.

[0249] Optionally, the first marker is located at the 5' end of the sequencing adapter in the sequencing read, and the second marker is located at the 3' end of the insert fragment in the sequencing read.

[0250] Optionally, the content of guanine (G) and cytosine (C) in the first marking code is 40% to 60%; and / or, the content of guanine (G) and cytosine (C) in the second marking code is 40% to 60%; and / or, the second marking code includes degenerate bases.

[0251] Optionally, the sequencing data processing apparatus provided in this embodiment further includes a generation module and a merging module.

[0252] Accordingly, the acquisition module 51 is further configured to, in response to receiving a sample tag generation request, randomly acquire an information bit code from a preset encoding library as the information bit code corresponding to the target sample, wherein the sample tag generation request includes the metadata of the target sample; the generation module is configured to generate a check bit code based on the information bit code using a preset RS algorithm; the merging module is configured to merge the information bit code and the check bit code to determine a first tag code; the generation module is further configured to generate a second tag code based on the first tag code and the metadata using a preset hash algorithm; and the determination module 52 is further configured to determine the first tag code and the second tag code as a tag for tagging the target sample.

[0253] Optionally, the generation module is further configured to use the fourth preset mapping rule in the preset RS algorithm to convert each base in the information bit encoding into a corresponding mathematical symbol; input each mathematical symbol into a preset polynomial set in the preset RS algorithm to calculate the check value; and convert the check value into the corresponding base based on the fourth preset mapping rule to generate the check bit encoding.

[0254] Optionally, the generation module is also used to generate a mixed number string based on the first marker code and metadata; calculate the hash value corresponding to the mixed number string using a preset hash algorithm; and convert the hash value into the corresponding base based on a third preset mapping rule to obtain the second marker code.

[0255] Optionally, the sequencing data processing apparatus provided in this embodiment further includes a deletion module.

[0256] Accordingly, the deletion module is used to delete the information bit encoding corresponding to the target sample from the preset encoding library.

[0257] Optionally, the storage module 53 is also used to associate the first tag code with metadata and store it in a preset metadata database.

[0258] Optionally, the metadata of the target sample includes: experimental batch number, timestamp, operator ID, and project number; the generation module is also used to regenerate a second tag code based on the first tag code and the metadata including the current timestamp in response to the determination that the timestamp has changed; the determination module 52 is also used to determine the first tag code and the regenerated second tag code as labels for tagging the target sample.

[0259] Figure 6 This is a schematic diagram of the structure of a sequencing data processing device provided in one embodiment of this application, as shown below. Figure 6 As shown, the sequencing data processing device 60 provided in this embodiment includes a processor 61 and a memory 62 that is communicatively connected to the processor.

[0260] The memory 62 stores computer-executable instructions; the processor 61 executes the computer-executable instructions stored in the memory 62 to implement the sequencing data processing method provided in the above embodiment. Related explanations can be understood by referring to the descriptions and effects corresponding to the steps in the accompanying drawings, and will not be elaborated upon here.

[0261] The program may include program code, which includes computer-executable instructions. Memory 62 may include high-speed RAM, and may also include non-volatile memory, such as at least one disk storage device.

[0262] In this embodiment, the processor 61 and the memory 62 are connected via a bus. The bus can be an Industry Standard Architecture (ISA) bus, a Peripheral Component Interconnect (PCI) bus, or an Extended Industry Standard Architecture (EISA) bus, etc. The bus can be divided into address bus, data bus, control bus, etc. For ease of representation, Figure 6 The bus is represented by a single thick line, but this does not mean that there is only one bus or one type of bus.

[0263] This application also provides a computer-readable storage medium storing computer-executable instructions. When the controller executes the computer-executable instructions, it implements the various steps in the methods described above.

[0264] This application also provides a computer program product, including a computer program that, when executed by a controller, implements the various steps in the methods described above.

[0265] The various embodiments described above in this application can be implemented in digital electronic circuit systems, integrated circuit systems, field-programmable gate arrays (FPGAs), application-specific integrated circuits (ASICs), application-specific standard parts (ASSPs), systems on chips (SOCs), complex programmable logic devices (CPLDs), computer hardware, firmware, software, and / or combinations thereof. These various embodiments may include implementations in one or more computer programs that can be executed and / or interpreted on a programmable system including at least one programmable processor, which may be a dedicated or general-purpose programmable processor, capable of receiving data and instructions from a storage system, at least one input device, and at least one output device, and transmitting data and instructions to the storage system, the at least one input device, and the at least one output device.

[0266] The computer-executable instructions used to implement the methods of this application may be written in any combination of one or more programming languages. These computer-executable instructions may be provided to the processor or controller of a general-purpose computer, special-purpose computer, or other programmable data processing device, such that when executed by the processor or controller, the computer-executable instructions cause the functions / operations specified in the flowcharts and / or block diagrams to be performed. The computer-executable instructions may be executed entirely on the machine, partially on the machine, partially on the machine and partially on a remote machine as a standalone software package, or entirely on a remote machine or electronic device.

[0267] In the context of this application, a computer-readable storage medium can be a tangible medium that may contain or store a program for use by or in conjunction with an instruction execution system, apparatus, or device. A computer-readable storage medium can be a machine-readable signal medium or a machine-readable storage medium. A computer-readable storage medium can be, but is not limited to, electronic, magnetic, optical, electromagnetic, infrared, or semiconductor systems, apparatus, or devices, or any suitable combination of the foregoing. More specific examples of computer-readable storage media include electrical connections based on one or more wires, portable computer disks, hard disks, random access memory (RAM), read-only memory (ROM), erasable programmable read-only memory (EPROM), optical fibers, compact disc read-only memory (CD-ROM), optical storage devices, magnetic storage devices, or any suitable combination of the foregoing. Alternatively, computer-readable storage media may include: resistive random access memory (RRAM), dynamic random access memory (DRAM), static random access memory (SRAM), enhanced dynamic random access memory (EDRAM), high-bandwidth memory (HBM), hybrid memory cube (HMC), and so on.

[0268] The systems and technologies described herein can be implemented in computing systems that include back-end components (e.g., as data electronic devices), or computing systems that include middleware components (e.g., application electronic devices), or computing systems that include front-end components (e.g., a user computer with a graphical user interface or web browser through which a user can interact with implementations of the systems and technologies described herein), or any combination of such back-end, middleware, or front-end components. The components of the system can be interconnected via digital data communication of any form or medium (e.g., a communication network). Examples of communication networks include local area networks (LANs), wide area networks (WANs), and the Internet.

[0269] It should be noted that, for the sake of simplicity, the foregoing method embodiments are all described as a series of actions. However, those skilled in the art should understand that this application is not limited to the described order of actions, as some steps can be performed in other orders or simultaneously according to this application. Secondly, those skilled in the art should also understand that the embodiments described in the specification are all optional embodiments, and the actions and modules involved are not necessarily essential to this application. In other words, the various forms of processes shown above can be used, with steps reordered, added, or deleted. For example, the steps disclosed in this application can be executed in parallel, sequentially, or in different orders, as long as the desired result of the technical solution disclosed in this application can be achieved, and this is not limited herein.

[0270] It should be further noted that although the steps in the flowchart are shown sequentially according to the arrows, these steps are not necessarily executed in the order indicated by the arrows. Unless explicitly stated herein, there is no strict order restriction on the execution of these steps, and they can be executed in other orders. Moreover, at least some steps in the flowchart may include multiple sub-steps or multiple stages. These sub-steps or stages are not necessarily completed at the same time, but can be executed at different times. The execution order of these sub-steps or stages is not necessarily sequential, but can be performed alternately or in turn with other steps or at least some of the sub-steps or stages of other steps.

[0271] It should be understood that the above-described device embodiments are merely illustrative, and the device of this application can also be implemented in other ways. For example, the division of units / modules in the above embodiments is only a logical functional division, and there may be other division methods in actual implementation. For example, multiple units, modules, or components may be combined, or integrated into another system, or some features may be ignored or not executed.

[0272] Furthermore, unless otherwise specified, the functional units / modules in the various embodiments of this application can be integrated into one unit / module, or each unit / module can exist physically separately, or two or more units / modules can be integrated together. The integrated units / modules described above can be implemented in hardware or as software program modules.

[0273] When an integrated unit / module is implemented in hardware, that hardware can be digital circuits, analog circuits, etc. The physical implementation of the hardware structure includes, but is not limited to, transistors, memristors, etc.

[0274] If the integrated unit / module is implemented as a software program module and sold or used as an independent product, it can be stored in a computer-readable storage device (CMD). Based on this understanding, the technical solution of this application, in essence, or the part that contributes to related technologies, or all or part of the technical solution, can be embodied in the form of a software product. This computer software product is stored in a memory and includes several instructions to cause a computer device (which may be a personal computer, server, or network device, etc.) to execute all or part of the steps of the methods of the various embodiments of this application. The aforementioned memory includes various media capable of storing computer-executable instructions, such as USB flash drives, read-only memory (ROM), random access memory (RAM), portable hard drives, magnetic disks, or optical disks.

[0275] In the above embodiments, the descriptions of each embodiment have their own emphasis. For parts not described in detail in a certain embodiment, please refer to the relevant descriptions of other embodiments. The technical features of the above embodiments can be combined arbitrarily. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as the combination of these technical features does not contradict each other, it should be considered within the scope of this specification.

[0276] Other embodiments of this application will readily occur to those skilled in the art upon consideration of the specification and practice of the invention disclosed herein. This application is intended to cover any variations, uses, or adaptations of this application that follow the general principles of this application and include common knowledge or customary techniques in the art not disclosed herein. The specification and examples are to be considered exemplary only.

[0277] It should be understood that this application is not limited to the precise structure described above and shown in the accompanying drawings, and various modifications and changes can be made without departing from its scope. Therefore, the specific embodiments described above do not constitute a limitation on the scope of protection of this application. Those skilled in the art should understand that various modifications, combinations, sub-combinations, and substitutions can be made according to design requirements and other factors. Any modifications, equivalent substitutions, and improvements made within the principles of this application should be included within the scope of protection of this application.

Claims

1. A method of processing sequencing data, characterized by, The method comprises: obtaining a sequencing read of a target sample, the sequencing read comprising a first marker code and a second marker code, the first marker code comprising: information bit encoding and check bit encoding, and the second marker code being a short sequence generated by a preset hash algorithm according to the first marker code and sample metadata; determining whether the information bit encoding is correct based on the check bit encoding; in response to determining that the information bit encoding is incorrect, determining the position of an error base in the information bit encoding and the corresponding correct base by using a preset Reed-Solomon RS algorithm to generate a corrected first marker code; obtaining metadata of the target sample from a preset metadata database based on the first marker code, the preset metadata database having stored therein metadata and first marker codes of a plurality of samples in association; determining whether the second marker code is correct based on the metadata and the corrected first marker code by using a preset hash algorithm; in response to determining that the second marker code is correct, determining the corrected first marker code and the second marker code as a label of the target sample, and storing the label and the metadata in association with the target sample.

2. The method of claim 1, wherein, The determination of whether the information bit encoding is correct based on the check bit encoding comprises: calculating a check value based on the information bit encoding by using a preset RS algorithm; matching the check value with the check bit encoding in the first marker code; if the matching is successful, it is determined that the information bit encoding in the first marker code is correct; if the matching fails, it is determined that the information bit encoding in the first marker code is incorrect.

3. The method of claim 2, wherein, The determination of the position of an error base in the information bit encoding and the corresponding correct base by using a preset Reed-Solomon RS algorithm to generate a corrected first marker code comprises: calculating an error base deviation value based on the check value and the check bit encoding in the first marker code; substituting the error base deviation value into a preset error position polynomial of a preset RS algorithm to calculate a polynomial result; determining the position of the error base according to a first preset mapping rule and the polynomial result; substituting the error base deviation value into a preset error amplitude polynomial of the preset RS algorithm to calculate an error amplitude; determining the correct base corresponding to the error base position according to a second preset mapping rule and the error amplitude; replacing the base at the error base position with the correct base to generate a corrected first marker code.

4. The method of claim 1, wherein, The determination of whether the second marker code is correct based on the metadata and the corrected first marker code by using a preset hash algorithm comprises: generating a mixed digital string according to the corrected first marker code and the metadata; calculating a hash value corresponding to the mixed digital string by using a preset hash algorithm; converting the hash value into a corresponding base based on a third preset mapping rule to obtain a second marker code corresponding to the corrected first marker code; matching the second marker code corresponding to the corrected first marker code with the second marker code in the sequencing read; in response to a successful matching, it is determined that the second marker code in the sequencing read is correct; in response to a failed matching, it is determined that the second marker code in the sequencing read is incorrect.

5. The method of claim 1, wherein, The first marker code is located at the 5' end of a sequencing adapter in the sequencing read, and the second marker code is located at the 3' end of an inserted fragment in the sequencing read.

6. The method of claim 1, wherein, The content of guanine (G) and cytosine (C) in the first marker code is 40% to 60%; And / or, the content of guanine (G) and cytosine (C) in the second marker code is 40% to 60%; And / or, the second marker code includes a degenerate base.

7. The method of claim 1, wherein, Before the sequencing data of the target sample is obtained, the method further comprises: In response to receiving a sample label generation request, randomly obtaining an information bit code from a preset encoding library as an information bit code corresponding to the target sample, the sample label generation request comprising metadata of the target sample; According to the information bit code, a check bit code is generated by using a preset RS algorithm, and the information bit code and the check bit code are combined to determine a first marker code; A second marker code is generated based on the first marker code and the metadata by using a preset hash algorithm; The first marker code and the second marker code are determined as a label for marking the target sample.

8. The method of claim 7, wherein, The method according to the information bit code, generating a check bit code by using a preset RS algorithm, comprises: Using a fourth preset mapping rule in the preset RS algorithm, each base in the information bit code is converted into a corresponding mathematical symbol; Each of the mathematical symbols is input into a preset polynomial group in the preset RS algorithm to calculate a check value; Based on the fourth preset mapping rule, the check value is converted into a corresponding base to generate the check bit code.

9. The method of claim 7, wherein, The method of generating a second marker code based on the first marker code and the metadata by using a preset hash algorithm comprises: Generating a mixed digital string according to the first marker code and the metadata; Calculating a hash value corresponding to the mixed digital string by using a preset hash algorithm; Based on a third preset mapping rule, the hash value is converted into a corresponding base to obtain the second marker code.

10. The method of claim 7, wherein, After the information bit code is randomly obtained from the preset encoding library as the information bit code corresponding to the target sample, the method further comprises: The information bit code corresponding to the target sample is deleted from the preset encoding library.

11. The method of claim 7, wherein, After the second marker code is generated, the method further comprises: The first marker code and the metadata are associated and stored in the preset metadata database.

12. The method of claim 7, wherein, The metadata of the target sample comprises an experiment batch number, a timestamp, an operator ID and a project number, and the method further comprises: In response to determining that the timestamp has changed, a second marker code is re-generated based on the first marker code and metadata including the current timestamp by using a preset hash algorithm; The first marker code and the re-generated second marker code are determined as a label for marking the target sample.

13. A sequencing data processing apparatus, characterized by, The device comprises: An acquisition module is configured to acquire a sequencing read of a target sample, the sequencing read comprising a first marker code and a second marker code, the first marker code comprising an information bit code and a check bit code, and the second marker code being a short sequence generated according to the first marker code and sample metadata by using a preset hash algorithm; determining whether the information bit encoding is correct based on the check bit encoding; the determining module is further configured to, in response to determining that the information bit encoding is incorrect, determine a position of an incorrect base and a corresponding correct base in the information bit encoding based on a preset Reed-Solomon (RS) algorithm, to generate a revised first marker code; the obtaining module is further configured to obtain metadata of the target sample from a preset metadata database based on the first marker code, the metadata database having metadata and first marker codes of a plurality of samples stored in association; the determining module is further configured to determine whether the second marker code is correct based on the metadata and the revised first marker code using a preset hash algorithm; the determining module is further configured to, in response to determining that the second marker code is correct, determine the revised first marker code and the second marker code as a label of the target sample; a storage module configured to store the label and the metadata in association with the target sample.

14. A sequencing data processing device, characterized by, The device comprises a processor and a memory connected in communication with the processor; the memory is configured to store computer execution instructions; the processor is configured to execute the computer execution instructions stored in the memory to implement the method of any one of claims 1-12.

15. A computer-readable storage medium, characterized in that, The computer readable storage medium has computer execution instructions stored therein, and the computer execution instructions are configured to be executed by a processor to implement the method of any one of claims 1-12.

16. A computer program product comprising a computer program configured to implement the method of any one of claims 1-12 when executed by a processor.

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