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Liquid-phase hybrid capture enriching liquor for genome DNA sequencing library and hybrid method adopting liquid-phase hybrid capture enriching liquor

A DNA sequencing and genome sequencing technology, applied in the field of molecular biology, can solve the problems of undisclosed formula and high price, and achieve the effect of shortening hybridization time, low cost and optimizing enrichment effect.

Inactive Publication Date: 2014-12-03
GENESEEQ TECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its hybridization solutions and hybridization methods are mostly aimed at the capture probes produced by our company. The length of the capture probes ranges from 60 to 150 bases, and the formula is not disclosed, so the price is expensive.

Method used

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  • Liquid-phase hybrid capture enriching liquor for genome DNA sequencing library and hybrid method adopting liquid-phase hybrid capture enriching liquor
  • Liquid-phase hybrid capture enriching liquor for genome DNA sequencing library and hybrid method adopting liquid-phase hybrid capture enriching liquor
  • Liquid-phase hybrid capture enriching liquor for genome DNA sequencing library and hybrid method adopting liquid-phase hybrid capture enriching liquor

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Embodiment 1

[0039] Genomic DNA sequencing library hybridization trap enrichment solution is prepared, which is composed of sodium phosphate buffer, sodium citrate buffer (SSC), SDS, EDTA and Denhardt solution. It can be stored for a long time at -20°C. The specific formula of its 2X concentration solution is as follows:

[0040] 0.5M Sodium Phosphate Buffer, pH 7.0 (1)

[0041] 1% SDS

[0042] 2mM EDTA

[0043] 2X SSC, pH7.0 (2)

[0044] 4X Denhardt's solution (3)

[0045] (1) Formula of 1M sodium phosphate buffer (pH7.0): 57.7ml1M Na 2 HPO 4 with 42.3ml 1M NaH 2 PO 4 mix

[0046] (2) 20X SSC formula: 3M NaCl, 300mM sodium citrate; adjust the pH value to 7.0.

[0047] (3) 50X Denhardt's solution: Denhardt's solution is a mixture of blocking reagents for hybridization. The solution contained 1% polysucrose (Ficoll, type 400), 1% polyvinylpyrrolidone, and 1% bovine serum albumin (BSA).

[0048] The above-mentioned hybridization solution carries out the method for hybridization,...

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Abstract

The invention discloses a liquid-phase hybrid capture enriching liquor for a genome DNA sequencing library and a hybrid method adopting the liquid-phase hybrid capture enriching liquor. The hybrid enriching liquor comprises phosphate buffer, sodium citrate buffer (SSC), SDS (lauryl sodium sulfate), EDTA (ethylene diamine tetraacetic acid) and Denhardt liquor. The hybrid liquor and the corresponding hybrid method are low in cost, simple and convenient to implement; the hybrid time can be effectively shortened, the needed DNA fragment can be precisely extracted, influences of a background value on a signal value can be reduced, and the hybrid target enriching effects can be optimized.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a formulation and a hybridization method of a genomic DNA sequencing library liquid-phase hybridization trapping enrichment solution. Background technique [0002] In the past few years, DNA sequencing technology has undergone a fundamental shift, from traditional Sanger sequencing to the so-called "Next Generation Sequencing" technology (Next Generation Sequencing, NGS). Its outstanding feature is massively parallel sequencing. NGS technology allows millions or even hundreds of millions of sequencing reactions to be performed simultaneously, thereby achieving a huge increase in sequencing throughput. The NGS technology platform needs to construct a fragment library suitable for massively parallel sequencing. The process of constructing a sequencing library involves DNA fragmentation, subsequent DNA repair and end processing (blunt ends or A overhangs). Finally, th...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12Q1/68
Inventor 邵华武邵阳
Owner GENESEEQ TECH INC
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