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Library building connector and method suitable for simplified genome sequencing of DNBSEQ technology

A technology of genome sequencing and technology, applied in the field of bioinformatics, can solve problems such as the complexity of database construction methods, and achieve the effect of simplifying genome sequencing and high sequencing quality

Pending Publication Date: 2022-08-05
CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with ordinary whole genome sequencing, genetic information covering the whole genome can be obtained at a lower cost; correspondingly, the method of building a library is more complicated

Method used

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  • Library building connector and method suitable for simplified genome sequencing of DNBSEQ technology
  • Library building connector and method suitable for simplified genome sequencing of DNBSEQ technology
  • Library building connector and method suitable for simplified genome sequencing of DNBSEQ technology

Examples

Experimental program
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Effect test

Embodiment 1

[0034] This example designs a linker for library construction, the linker is composed of one or several pairs of DNA double-stranded fragments, and each pair of linkers is divided into an upstream linker sequence (F) and a downstream linker sequence (R).

[0035] like figure 1 As shown, the upstream linker F contains the following structure:

[0036] (1) An enzyme cutting end portion (E1) used for complementary pairing with a sticky end produced by a restriction endonuclease, and the E1 sequence can be replaced according to the selected restriction endonuclease, such as a linker sequence. E1 is a sticky end, which contains two DNA single strands, forward strand E1-A and reverse complementary strand E1-B. Among them, E1-A from 5' to 3' direction is the double-stranded partial sequence in the sticky end left after the corresponding restriction endonuclease digests DNA; E1-B from 3' to 5' direction is the restriction endonuclease The reverse strand sequence left after enzymatic...

Embodiment 2

[0052] This example uses the sequencing adapter shown in Example 1 to design a simplified genome sequencing method suitable for DNBSEQ technology. The specific process is as follows:

[0053] 1. Adapt to different restriction enzymes and Barcode / Index

[0054] According to the linker structure and general formula designed in this invention, the linker structure can be designed for the following sequencing requirements:

[0055] (1) When genomic DNA was digested with EcoRI-MspI double enzyme, library construction was performed for 3 individuals. 3 kinds of Index, 3 kinds of Barcode-1, 3 kinds of Barcode-2 are needed in order to meet the sequencing requirements, and the E1 and E2 fragments are respectively replaced by the enzyme cut sticky ends of EcoRI and MspI, specifically as shown in Table 2 (SEQ ID NO.7- 18):

[0056] Table 1: When using EcoRI-MspI double restriction enzyme to digest genomic DNA, 3 individual Barcodes, Index and restriction sites

[0057]

[0058] ...

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Abstract

The invention relates to the technical field of biological information, in particular to a library building connector and method suitable for simplified genome sequencing of a DNBSEQ technology. The linker comprises an upstream linker sequence and a downstream linker sequence; the upstream linker sequence comprises an upstream Barcode fragment, a Q1-1 fragment, an Index fragment and a Q1-2 fragment which are connected in sequence; the downstream linker sequence comprises a downstream Barcode fragment and a Q2 fragment; wherein the upstream linker sequence is reversely complementary, and the Q2 fragment part in the downstream linker sequence is reversely complementary; the upstream Barcode fragment and the downstream Barcode fragment are not equal in length. The invention provides a library building joint and designs a simplified genome sequencing method suitable for a DNBSEQ technology based on the library building joint, and genome sequencing can be accurately, quickly and effectively carried out.

Description

technical field [0001] The invention relates to the technical field of biological information, in particular to a library-building linker and method suitable for simplified genome sequencing of DNBSEQ technology. Background technique [0002] DNA sequencing technology is the technology to determine the sequence of DNA molecules. Library construction technology, that is, library construction, refers to the repair of DNA existing in samples such as blood, body fluids, tissues or feces and ligated to a specific DNA fragment, that is, a linker sequence, so that it can be sequenced on a specific technology platform. process. As an essential step for sequencing, library building has a variety of technical steps according to different application requirements. Correspondingly, the adapters also have different structures and sequences. [0003] Sequencing technology is an important foundation for many research fields such as genetics and molecular biology in biological research to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806C12N15/10C12N15/11C40B50/06C40B80/00
CPCC12Q1/6806C12N15/1093C40B50/06C40B80/00C12Q2521/301C12Q2525/191
Inventor 胡晓湘边成谈成李宁
Owner CHINA AGRI UNIV
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