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Sequencing library adapters with increased stability

A sequencing library and double-stranded connector technology, applied in the field of sequencing library connectors, can solve problems affecting detection accuracy and specificity, low connection efficiency, unfavorable gene detection, etc., achieve excellent connection efficiency, meet needs, and reduce adapter dimerization body's effect

Active Publication Date: 2021-11-09
上海英基生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are few adapters used in conjunction with the BGI sequencing platform, and most of the adapters that have appeared in the market have low connection efficiency, which is not conducive to the genetic detection of some samples with low content, affecting the accuracy and specificity of the detection. It is urgent to develop a high-connection efficient connector

Method used

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  • Sequencing library adapters with increased stability
  • Sequencing library adapters with increased stability
  • Sequencing library adapters with increased stability

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Connector Construction Library Test

[0027] First, ECOLI GDNA sample preparation

[0028] Take the Escherichia coli liquid, select the bacterial genomic extract kit to extract ECOLI GDNA according to the specification. After the extraction, the library is put into 50 ng. The joint uses B1 (B1 as a double-link, B1R1 sequence such as SEQ No. 2), B2 (B2 is a double link, B2 R1 sequence such as SEQ No.3 , B2R2 sequences such as SEQ No. 4), B3 (B3 is a double-link joint, B3R1 sequence such as SEQ No. 5, B3R2 sequence such as SEQ No. 6) Each group of experiments two repetitions, and use Illumina short connections (such as Figure 5 The indicated as a control.

[0029] Second, DNA library construction

[0030] In this embodiment, the DNA library is constructed using ABClonal commercial Mega With Ddres DNA lib preprum for MGI (RK20241). The specific library preparation procedure is detailed in the Commercial Kit Operating Instructions. The following is the basic DNA libr...

Embodiment 2

[0054] Example 2: Different templates of different templates

[0055] First, Human GDNA, FFPE sample preparation

[0056] Take some sample organization, select the appropriate genomic extract kit to extract the Human GDNA according to the specification. The FFPE sample was extracted with xylene extraction samples. After the extraction, the mechanical interrupt concentration was 24.6 ng / ul; 19 ng / ul, 150 bp, respectively. Two repetitions were repeated for 50 ng, 10 ng, and 1 ng of each group.

[0057] Second, DNA library construction

[0058](1) The specific construction library process is the same as in Example 1

[0059] (2) The joint concentration is 15μm

[0060] (3) The number of cycles is performed according to the following table

[0061]

[0062] Third, the library quantification

[0063] Quantitatively quantitatively using Qubit, quantitative results are as follows:

[0064]

[0065]

[0066]

[0067] B1, B2, B3 joints in this design are small in the 50 ng inpu...

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Abstract

The present invention discloses a sequencing library linker with increased stability. Using the sequencing library adapter of the present invention, the yield of library construction is better than or equal to that of other sequencing platforms; the sequencing library adapter connection efficiency of the present invention is excellent, which can reduce the generation of adapter dimers, so as to meet the needs of scientific research units and enterprises for BGI. The need for a sequencing platform.

Description

Technical field [0001] The present invention relates to the field of biotechnology, and specifically refers to a sequencing library joint having a stability increase. Background technique [0002] The earliest sequencing method is a chemical degradation method for the final sequencing method of Sanger inventions in 1977 and the chemical degradation of the same year A.MAXAM and W.Gilbert. Among them, the Sanger method has reached 1000bp, accuracy is 99.999%, but the shortcomings of high sequencing cost, low flux, etc., have not fully met the needs of research. With the continuous development of science and technology, people's continuous improvement and development of sequencing methods, the second generation sequencing came into being. The new generation of sequencing has rapidly occupying most of the market in the sequencing industry, which has rapidly occupying most of the sequencing industries, which is based on countries such as Roche, Illumina and other foreign sequencing pl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C40B80/00
CPCC40B80/00
Inventor 冯延叶柴智张会赖煦卉孙大鹏
Owner 上海英基生物科技有限公司
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