18 results about "GC-content" patented technology

The application relates to a sarcoma fusion gene nanopore sequencing primer set and application thereof, wherein the primer set comprises five pairs of specific primers (Primer 1-5) with fixed sequences, each primer follows the design principle of 18-25 bp in length, 40%-60% in GC content and strict matching of a target sequence at a 3' end, and the length of an amplification product is 300-800 bp, which is suitable for nanopore sequencing. Through qPCR verification, the primer set has single-peak melting curves, close-to-100% amplification efficiency, strong specificity and excellent amplification performance.

A microbial gene editing scheme automatic design method, system and storage medium; the method comprises: selecting a design object for different types of strains, the design object is bacteria or fungi; after determining the target gene, obtaining the target gene information from the NCBI database; the design scheme of bacteria comprises the following steps: determining the gRNA design region; selecting the optimal gRNA sequence; doing sequence complexity and GC content analysis; the design scheme of fungi uses a frameshift scheme: the gRNA of the frameshift scheme is set at any position not overlapping with other genes after the start codon in the gene; select gRNA sequence in gRNA design region, select the optimal gRNA sequence; design two pairs of four primer sequences for homologous arm identification primer; this scheme can replace the complicated and time-consuming manual scheme, save a lot of time, solve the problem of high cost in the design of the best site and the design of sgRNA sequence scheme of artificial gene knockout.

This invention discloses a method for constructing a multi-target amplicon abundance standard for microorganisms and its application, belonging to the fields of molecular biology detection and microbiome analysis. This standard material screens sequences of common human gut microbiota strains, designs and adds universal primers, and obtains 14 DNA fragments of different lengths through gene synthesis, cloning, and purification, which are then mixed according to a preset abundance. It retains natural characteristics such as primer binding sites and GC content, and can systematically correct technical deviations in amplicon sequencing, solving the problem of species abundance distortion. This standard material functions as both a non-homologous internal reference and a homologous external reference, and can be used for laboratory proficiency testing, reagent kit performance evaluation, and cross-platform data calibration, promoting the leap from relative qualitative to absolute quantitative research in microbiome studies and providing metrological support for the standardization and precision of detection results.

The invention belongs to the technical field of biology, and particularly relates to a DNA polymerase mutant for improving amplification efficiency of a high GC sample and application of the DNA polymerase mutant. According to the invention, DNA polymerase mutants KODG and KODC are obtained through site-specific mutagenesis, and on the basis of retaining the capability of amplifying a DNA template with conventional GC content, the capacity of amplifying a DNA template with high GC content (gt; compared with the prior art, the method has the advantages that the amplification efficiency of a DNA template (3kb, 60%) is improved, effective amplification of difficult-to-amplify fragments with 3kb, 95% GC content and the like is successfully realized, and the technical bottleneck of amplification failure caused by high thermal stability and difficulty in complete melting of the existing polymerase in a high-GC region is overcome, so that the application range of the DNA polymerase in complex template amplification is expanded, and the application prospect is broad. And a more reliable enzymology tool is provided for related detection and identification of high-GC-content samples.

The invention discloses a method and a system for realizing DNA storage by aiming at multi-rule rotation coding of Chinese texts, and relates to the technical field of DNA storage. Encoding the Chinese text by using a five-stroke font input method; mapping is carried out according to the positive and negative code tables, and an interval balance and dynamic detection strategy is introduced in the mapping process to control GC content distribution in intervals and the probability of occurrence of homopolymers between the intervals. Scattering and recombining the sequence by using block coding, and compressing by using RLE coding, wherein the RLE coding generates a sequence file and a run-length file; performing GC content constraint on the sequence file by using cross coding; and br compression is adopted for the run-length file to further improve the compression ratio. And generating a DNA sequence through rotary coding. According to the method, five-stroke coding, positive and negative code table mapping and sequence reconstruction strategies are introduced, so that the information storage density is higher, the local GC content of the DNA sequence is more stable, the homopolymer length is smaller, the unexpected motif proportion is lower, and the data has higher reliability and safety in the storage process.

The present application relates to the field of DNA methylation modification detection, and particularly relates to nucleic acid sequence molecules and DNA methylation detection methods. The nucleic acid sequence molecules comprise at least a probe sequence, the probe sequence comprises at least one CpG site, at least one of which is a to-be-detected CpG site; the probe sequence comprises a methylation probe sequence and a non-methylation probe sequence, and the two probe sequences are modified with different fluorescent groups; and the probe sequence at least meets one of the following conditions: i) within the region of the to-be-detected CpG site and the 2 nt upstream and downstream of the to-be-detected CpG site, the overall GC content of the methylation probe is not more than 50%; ii) within the region of the to-be-detected CpG site and the 2 nt upstream and downstream of the to-be-detected CpG site, the overall GC content of the methylation probe is more than 50%, and the to-be-detected CpG site in the probe sequence is modified with LNA. The method of the present application can significantly improve the specificity and sensitivity of CpG site methylation detection.

The invention discloses a stable and high-efficiency high-fidelity DNA polymerase mutant and a preparation method thereof, relates to the fields of genetic engineering and minute biology, and aims to modify a high-fidelity DNA polymerase by rationally designing mutation sites and optimizing additives, enhance the thermal stability, extension efficiency and continuous synthesis capability of the high-fidelity DNA polymerase and maintain the high-fidelity characteristic of the high-fidelity DNA polymerase. The optimized DNA polymerase shows significantly improved amplification speed in the PCR amplification process, can efficiently synthesize longer DNA fragments, and is suitable for amplification of complex templates (such as sequences with high GC content or rich secondary structures). According to the method, the synthesis efficiency and accuracy of the long-fragment DNA are remarkably improved, and a more efficient tool is provided for genome sequencing, gene cloning, in-vitro diagnosis and the like.

The present application relates to a kind of primer group, kit and method for detecting ATXN8OS and ATXN8 gene (CTA·TAG) n And (CTG·CAG) n Trinucleotide repeat number, belong to gene detection technical field.The primer group includes first primer pair and second primer pair;The forward primer sequence of the first primer pair is as shown in SEQ ID NO.1, and the reverse primer sequence is as shown in SEQ ID NO.2;The forward primer sequence of the second primer pair is as shown in SEQ ID NO.1, and the reverse primer sequence is as shown in SEQ ID NO.3.The present application is realized by the design of two primer pairs, complementary verification is realized, detection result is comprehensive and reliable, make up the technical deficiency of one generation sequencing and two generation sequencing in long fragment, high GC content repeat sequence detection, and detection cost is low, detection cycle is short.

The application discloses a PTPRQ gene mutation site related to non-syndrome deafness, the mutation site is a novel pathogenic mutation site on a human PTPRQ gene, and is one or more combinations selected from c.4723C>T, c.6040C>T, c.1811T>C, c.5687-6del and c.6560C>T. A specific probe set for detecting a PTPRQ gene mutation, the probe set is a double-stranded DNA probe, is designed according to a PTPRQ gene 45-exon sequence of a transcript NM_001145026 and a flanking region of each 5' end and 3' end of each exon with an extension of 50 bp, contains 169 probes in total, the length of a single probe is 120 bp, a shingled design is adopted, there is an overlapping region between adjacent probes, and the probe density is dynamically adjusted according to the GC content of a target sequence. A detection method of a PTPRQ gene mutation, with peripheral blood or a blood sheet of a to-be-tested person as a sample, extracting genomic DNA, performing targeted capture enrichment by using the specific probe set, combining high-throughput sequencing with standardized bioinformatics analysis, and judging whether the PTPRQ gene mutation exists in the to-be-tested sample.

The application belongs to the technical field of fluorescent quantitative PCR, and provides an enzyme digestion-fluorescent quantitative PCR detection method. The method comprises the following steps: step one, configuring a reaction mixture, wherein the reaction mixture comprises amplification reagents, a restriction endonuclease, Taq DNA polymerase, UDG enzyme, primers and a probe; and step two, setting a reaction program and a reaction system. Non-target templates in a qPCR reaction system are degraded by enzyme digestion of a restriction endonuclease, thereby reducing non-specific amplification; the proportion of target templates is increased, thereby avoiding the interference of non-target templates and improving detection sensitivity and detection stability. In particular, the method can effectively detect high GC content templates, such as TERT gene promoters or CpG island methylation.

The invention discloses a universal insertion sequence and application thereof in preparation of multiple PCR template nucleic acid. The invention provides a universal insertion sequence and a method for preparing multiple PCR template nucleic acid based on the universal insertion sequence, and the method comprises the following steps: firstly, designing universal insertion sequences with different GC contents, and distributing a universal insertion sequence with similar GC content for each multiple PCR target amplicon; the method comprises the following steps: firstly, synthesizing a universal insertion sequence primer, then synthesizing a hybrid PCR primer containing the universal insertion sequence primer and a multiplex PCR target amplicon primer, and introducing the multiplex PCR target amplicon into the universal insertion sequence through one-step PCR amplification, so as to obtain a set of artificial chimeric DNA template containing the multiplex PCR primer, and the artificial chimeric DNA template can be used as multiplex PCR template nucleic acid for developing and optimizing a multiplex PCR system.

The present invention relates to a Klotho messenger RNA (mRNA), wherein the mRNA has a 5' CAP region, a 5' untranslated region (5'-UTR), a coding region encoding a Klotho polypeptide, a 3' untranslated region (3'-UTR), and a polyadenosine tail (poly-A tail), wherein the Klotho polypeptide comprises the human Klotho KL1 domain, preferably wherein the coding region encoding the Klotho polypeptide comprises an RNA sequence having at least 80% sequence identity with SEQ ID NO: 1, and wherein the coding region encoding the Klotho polypeptide preferably has a GC content of at least 54%.

The application discloses a quaternary DNA storage method and system based on entropy coding and rotation constraint coding, a storage medium and a terminal. First, compared with traditional binary coding, quaternary coding can improve storage density, thereby storing more data in smaller space. Second, dynamic rotation coding ensures that the DNA sequence meets strict biochemical constraints, including homopolymer length limitation, GC content balance and avoidance of specific harmful sequences, optimizes the synthesis and sequencing process of DNA, and reduces the error rate. In addition, the use of tANS coding provides higher data compression efficiency, further reducing storage costs. These technical innovations not only improve the performance of large-scale data storage, but also enhance the security and accuracy of data decoding, ensuring the integrity of data after long-term storage.

The present invention provides Klotho messenger RNA (mRNA) having a 5' CAP region, a 5' untranslated region (5'-UTR), a coding region encoding a Klotho polypeptide, a 3' untranslated region (3'-UTR), and a polyadenosine tail (poly-A tail), wherein the Klotho polypeptide comprises the KL1 domain of human Klotho, preferably the coding region encoding the Klotho polypeptide comprises an RNA sequence having at least 80% sequence identity with SEQ ID NO: 1, and preferably the coding region encoding the Klotho polypeptide has at least 54% GC content.

By means of a method, a genome editing technique can be provided which is subject to a general organism, in particular actinomycetes, in which the engineered organism or cell does not conform to a recombinant and can avoid an off-target effect, in which the engineered organism or cell does not conform to the recombinant. The present invention relates to a method for deleting, substituting or inserting a target base sequence in a region to be subjected to genome editing in a living being or a cell to be subjected to genome editing, the method comprising: (1) a step for introducing a type II restriction endonuclease recognition sequence into the region to be subjected to genome editing; and (2) a step for causing a double-strand break (DSB) to occur in a genome by expressing in the organism or cell a type II restriction endonuclease that recognizes the introduced type II restriction endonuclease recognition sequence, after the steps (1) and (2) are performed, in the organism or cell, a sequence of the type II restriction endonuclease recognition sequence is introduced into the organism or cell, and the sequence of the type II restriction endonuclease recognition sequence is converted into a sequence of the type II restriction endonuclease recognition sequence into a sequence of the type II restriction endonuclease recognition sequence. The subject region is edited by homologous recombination repair with a gene sequence for genome editing, and the GC content in the genome in the organism or cell is 65% or more.

The invention discloses a probe set, a kit and a method for detecting blood tumor genes, the probe set comprises a plurality of probes, 1 / 3-2 / 3 of the two adjacent probes are overlapped during targeted detection, and each basic group in a targeted detection area is covered by the two probes. The invention provides a 2 * Tiling designed probe group, and each basic group in a detection area is covered by two primers, so that the capture efficiency is improved by about 20% compared with a conventional imbricated designed probe. For fusion detection, not only common driver genes but also partner genes are contained. Probe design is carried out in a mode of laying probes at a breakpoint and an exon boundary region, and the detection sensitivity of fusion variation is remarkably improved. In addition, the probe density is adjusted according to the GC content, and the high-GC region adopts a'high-density coverage 'strategy, so that the coverage degree is remarkably improved. Aiming at the characteristic that the blood tumor fusion partner gene is highly homologous, the optimized sequence is used as a reference sequence to detect the IGH / IGJ / IGK region, so that the problem of multiple comparison is avoided.

The application belongs to the field of gene sequencing, and relates to a standard substance for calibrating a high-throughput sequencer, a preparation method and application thereof. 97 DNA fragments of 320 bp in length are selected from a genome. The DNA fragments are optimized through the principle of chirality, reverse design, GC content homogenization, structural stability control, series modularization integration, and are added with tags and primers at two ends. The candidate fragment library with a length of 456 bp is prepared through gene fragment synthesis, cloning and identification, and target fragment amplification and purification. After correction, the above-mentioned fragments are mixed according to the correction ratio to obtain a multi-fragment DNA mixed standard substance. Nucleic acid quantification, sequencing, GC content calculation, uniformity test and stability research are carried out on the standard substance, and value assignment and uncertainty evaluation are completed, so that the metrological traceability is ensured. The high-throughput sequencer calibration standard substance containing 97 456 bp fragments is successfully developed, and the measurement characteristics are systematically confirmed.

This invention discloses the identification, construction, and application of plant centromere tandem repeat sequences. The method includes: obtaining the target plant genome sequence; detecting tandem repeats through prefix sum vectorization to obtain candidate repeat regions and periods; extracting repeat units as candidate monomers and eliminating low-complexity noise; calculating a multi-feature fusion score for candidate monomers, the score including period consistency, genome enrichment, sequence complexity, and GC content shift, and screening based on the score; clustering the screened candidate monomers to obtain monomer subtypes; performing phase correction on monomers within subtypes; and constructing a consensus sequence based on the corrected sequence. This application achieves quantitative evaluation of centromere attribution probability through multi-feature fusion scoring, combined with exhaustive cyclic shift phase correction, to obtain a consensus sequence without relying on external tools or known motifs. Taking the Arabidopsis Col-CEN genome as an example, the optimal period is 178 bp, the column consistency rate is 0.9272, and the alignment consistency with the reference sequence exceeds 98%.