A kind of construction method and application of dna large fragment library

A construction method and large fragment technology, applied in the field of molecular biology, can solve the problems of low effective data rate and short read length, and achieve the effect of improving the effective data rate

Active Publication Date: 2022-05-17
ZHEJIANG ANNOROAD BIO TECH CO LTD +2
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of next-generation sequencing is the short read length. The Roche 454, which has the longest read length in the second-generation sequencer, can only perform DNA sequencing of up to 400 bp. According to the overlapping sequences on different small fragments, their sequencing structures are spliced ​​into contigs of different sizes (fragment contigs)
However, the effective data rate of the large fragment library obtained by the existing method is low after sequencing

Method used

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  • A kind of construction method and application of dna large fragment library
  • A kind of construction method and application of dna large fragment library
  • A kind of construction method and application of dna large fragment library

Examples

Experimental program
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Effect test

Embodiment 1D

[0073] Example 1 DNA Large Fragment Library Construction

[0074] 1. Large fragment processing of DNA samples

[0075] 1.1 Take 2 mL of whole blood from healthy people and extract it according to the operating instructions of the blood genome DNA extraction system (0.1-20 mL) (Tiangen Biochemical Technology Co., Ltd.) to obtain DNA samples. Take 10 μg of DNA sample, use the HydroShear Plus DNA Fragmentation Instrument to fragment large fragments of DNA samples according to the instructions, and the target large fragment length is 10kb.

[0076] 1.2 Prepare 0.8% agarose gel, use TIANGEN company 1Kb DNA Ladder as the molecular weight standard, and electrophoresis at 100V for 2 hours. After electrophoresis, the gel was taken out and stained in TAE containing EB dye for 20 minutes. Fragments around 9-12kb were cut and glued under ultraviolet irradiation.

[0077] 1.3 Put the excised gel into a clean 2.0mL centrifuge tube that has been weighed, and use the agarose gel extractio...

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Abstract

The invention relates to a method for constructing a large DNA fragment library and its application. The method for constructing a large DNA fragment library provided by the present invention is used for the construction of a large DNA fragment library. By using the FLP / FRT specific recombination system to optimize the library construction process, the effective data rate of the sequencing results of the large fragment library is improved.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a method for constructing a DNA large fragment library, a DNA large fragment library constructed by the method and an application of the DNA large fragment library in sequencing. Background technique [0002] For a species whose genome sequence is unknown or has no genome information of related species, its genome DNA fragments of different lengths and their libraries are sequenced, and then spliced, assembled and annotated by bioinformatics methods to obtain the complete genome of the species Sequence map, called genome de novo sequencing, also called de novo sequencing. Today, with the rapid development of genomics, the combination of de novo sequencing and comparative genomic methods can be used to explore the origin and evolution of the species, and to study the molecular mechanisms of its growth and development, shape production and environmental adaptation, whic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10C12Q1/6806C12Q1/6869C40B50/06
CPCC12N15/1093C12Q1/6806C12Q1/6869C40B50/06C12Q2531/113C12Q2525/191C12Q2521/507C12Q2535/122
Inventor 梁峻彬李小林张介中韩典昂玄兆伶李大为陈重建
Owner ZHEJIANG ANNOROAD BIO TECH CO LTD
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