A method for identifying sequencing samples and its application

A sequencing and sample technology, applied in the field of high-throughput sequencing, can solve the problem of high cost, achieve the effect of simple method, large number of numbers, and qualitative and quantitative analysis

Active Publication Date: 2021-09-24
江西海普洛斯生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the sequencing adapter of this invention is prepared by chemical synthesis, which is expensive and still has a false positive rate of about 0.005%.

Method used

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  • A method for identifying sequencing samples and its application
  • A method for identifying sequencing samples and its application
  • A method for identifying sequencing samples and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] figure 1 Shown is a flowchart of a method for analyzing a sequenced sample, the method comprising the following steps:

[0046] Step 110, constructing a DNA tag library, the homology of the DNA tag library and the sequencing sample library is 5%;

[0047] Step 120, generating the binary identification code number of the sequencing sample, and determining the identification sequence according to the binary identification code;

[0048] Step 130, the identification sequence is mixed with the sequencing sample;

[0049] Step 140, library construction and sequencing;

[0050] Step 150, identification sequence reading and quality control analysis;

[0051] Step 160, if the identification sequence is the same as the original code, then there is no pollution between the samples, and the quality inspection is passed; if the identification sequence is different from the original code, then there is pollution between the samples, and the pollution source analysis and quantificat...

Embodiment 2

[0052] Example 2 Construction of the Identification Sequence

[0053] (1) The sequencing sample library contains 256 human sequencing samples, and the sequencing samples are numbered with an 8-digit binary identification code;

[0054] (2) Construct the DNA tag library shown in Table 1, and the homology between the DNA tag library and the sample is 5%.

[0055] Table 1 contains the DNA tag library of 8 120bp DNA sequences

[0056]

[0057]

[0058]

Embodiment 3

[0059] Example 3 Construction of library and high-throughput sequencing

[0060] In this example, the library construction is performed on the cfDNA sample with ID 85, such as figure 2 As shown, the binary identification code of the human-derived sequencing sample with ID 85 is 01010101.

[0061] (1) Add Tag 1, Tag 3, Tag 5, and Tag 7 to 30ng cfDNA sample in an amount of 3×10 -4 ng, mix evenly, and carry out library construction;

[0062] (2) After the cfDNA is end-repaired, the 3' end is subjected to an A-tail ligation reaction, and the sequencing adapter is subjected to a T-tail ligation reaction, and the sequencing adapter is connected to the target cfDNA fragment by the principle of complementary base pairing;

[0063] (3) Use the pan-cancer specific probes independently developed by Shenzhen Hypros Biotechnology Co., Ltd. to capture related genes and tag library DNA sequences. The ends of the probes are modified with biotin. After the DNA and the probe are hybridized ...

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Abstract

The present invention provides a method for identifying a sequencing sample and its application. The method comprises the following steps: (1) constructing an injective relationship between a sequencing sample library and a DNA tag library; (2) according to the injective relationship described in step (1), relationship, using the DNA tags in the DNA tag library to identify the sequencing samples. The invention determines the DNA tag combination uniquely corresponding to the sequencing sample by constructing the injective relationship between the sequencing sample library and the DNA tag library, and different sequencing samples correspond to different DNA tag combinations, realizing the specificity of the DNA tag to the sequencing sample logo.

Description

technical field [0001] The invention belongs to the technical field of high-throughput sequencing, and relates to a method for marking a sequencing sample and an application thereof. Background technique [0002] Next generation sequencing technology (next generation sequencing, NGS) has a wide range of applications in clinical and scientific research fields. This technology has the characteristics of high throughput. However, due to the limited amount of sequencing data of a single sample, multiple samples are usually mixed and put on the machine at the same time. sequencing. In order to distinguish different samples, 6-8 different index base sequence adapters need to be added to different samples during the library construction process. [0003] Although this method is simple and feasible, it is prone to contamination between samples and errors. The main reasons include the following aspects: First, the index sequences between different samples generally only have 2-3 di...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G16B30/10G16B50/00C12Q1/6869
CPCC12Q1/6869C12Q2535/122C12Q2537/165
Inventor 陈实富刘明张晓妮许明炎卢超
Owner 江西海普洛斯生物科技有限公司
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