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Sequencing library linker with increased stability

A sequencing library and double-stranded adapter technology, applied in the field of sequencing library adapters, can solve the problems of unfavorable gene detection, affecting detection accuracy and specificity, low connection efficiency, etc., achieve excellent connection efficiency and reduce the generation of adapter dimers , to meet the desired effect

Active Publication Date: 2020-05-12
上海英基生物科技有限公司
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  • Abstract
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  • 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 linker with increased stability
  • Sequencing library linker with increased stability
  • Sequencing library linker with increased stability

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Joint library building test

[0027] 1. Ecoli gDNA sample preparation

[0028] Take the Escherichia coli liquid, select the bacterial genome extraction kit to extract Ecoli gDNA and extract according to the instructions. After extraction, put 50ng into the library, use B1 (B1 is a double-stranded linker, B1R1 sequence is as SEQ No.1, B1R2 sequence is as SEQ No.2), B2 (B2 is a double-stranded linker, B2 R1 sequence is as SEQ No.3) , B2R2 sequence such as SEQ No.4), B3 (B3 is a double-stranded linker, B3R1 sequence such as SEQ No.5, B3R2 sequence such as SEQ No.6) each group of experiments is repeated twice, and Illumina short linker (such as Figure 5 shown) for comparison.

[0029] 2. DNA library construction

[0030] In this example, ABclonal commercialized Mega with DDREs DNA Lib Prep Kit for MGI (RK20241) was used to construct a DNA library. For specific library preparation steps, please refer to the operating instructions of the commercial kit. The ba...

Embodiment 2

[0054] Example 2: Tests of different inputs on different templates of joints

[0055] 1. Human gDNA, FFPE sample preparation

[0056] Take human sample tissue, select the appropriate genome extraction kit to extract Human gDNA and extract according to the instructions. Take the paraffin-fixed samples, and use the xylene extraction method to extract the FFPE samples. After extraction, the concentrations after mechanical disruption were 24.6ng / ul; 19ng / ul, about 150bp. 50ng, 10ng, and 1ng were used to build a library, and each experiment was repeated twice.

[0057] 2. DNA library construction

[0058](1) The specific process of building a database is the same as the process of building a database in Example 1

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

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

[0061]

[0062] 3. Library quantification

[0063] Qubit was used to quantify the library, and the quantitative results are shown in t...

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Abstract

The invention discloses a sequencing library linker with increased stability. By adopting the sequencing library linker, the library construction yield is superior to or equal to that of linkers of other sequencing platforms. The sequencing library linker provided by the invention is excellent in linking efficiency, can reduce the generation of linker dimers, and can meet the requirements of scientific research institutions and enterprises on BGI sequencing platforms.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a sequencing library adapter with increased stability. Background technique [0002] The earliest sequencing method, that is, generation sequencing, was the terminal termination sequencing method invented by Sanger in 1977 and the chemical degradation method invented by A.M.Maxam and W.Gilbert in the same year. Among them, the read length of Sanger sequencing can reach 1000bp, and the accuracy is as high as 99.999%. However, the shortcomings of high sequencing cost and low throughput can no longer fully meet the needs of research. With the continuous development of science and technology, people continue to improve and develop sequencing methods, and second-generation sequencing came into being. Due to its advantages of lower cost, higher throughput, and faster speed, the next-generation sequencing has quickly occupied most of the sequencing industry market, with foreign sequencing ...

Claims

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Application Information

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Patent Type & Authority Applications(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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