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Kit and method for constructing DNA library

A DNA library and kit technology, applied in chemical library, combinatorial chemistry, library creation, etc., can solve the problem of low amplification efficiency of kits, achieve improved adapter connection efficiency, high amplification efficiency, and improved amplification efficiency Effect

Active Publication Date: 2016-05-25
BEIJING NOVOGENE TECH CO LTD
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Problems solved by technology

[0006] The main purpose of the present invention is to provide a kit and method for constructing a DNA library, to solve the problem of low amplification efficiency of kits in the prior art

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  • Kit and method for constructing DNA library

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Embodiment Construction

[0022] It should be noted that, in the case of no conflict, the embodiments in the present application and the features in the embodiments can be combined with each other. The present invention will be described in detail below with reference to the accompanying drawings and examples.

[0023] It should be noted that the various commercially available reagents described in the present invention do not include a complete set of kits for library construction in the existing market or a single reagent in the kits, but refer to reagents sold in a separate form.

[0024] In the present invention, M is the abbreviation of molar concentration mol / L, mM is the abbreviation of molar concentration mmol / L, and μM is the abbreviation of molar concentration μmol / L.

[0025] 10XT4 DNA ligase buffer is 700mmTris-HCl (PH8.0), 100mmMgCl2 and 50mmDTT; wherein, the working concentration of 1X means: 70mMTris-HCl (PH8.0), 10mMMgCl2 and 5mMDTT; the working concentration of 0.5X means: 35mM Tris-H...

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Abstract

The invention provides a kit and method for constructing a DNA library. The kit comprises an enzymic reagent, a buffer solution and a joint combination, wherein the enzymic reagent comprises a first enzyme mixed liquid, T4DNA ligase and a second enzyme mixed liquid, and the first enzyme mixed liquid is formed by mixing T4 Polynucleotide Kinase, T4DNA polymerase, a Klenow fragment and rTaq enzyme; the second enzyme mixed liquid is a 2X KAPA HiFi HotStart pre-mixed solution; the buffer solution comprises a first buffer solution and a second buffer solution, and the first buffer solution is formed by mixing a 10X T4DNA ligase buffer solution, dNTP mixed liquor and ATP; the second enzyme mixed liquid is formed by mixing ATP, Tris-HCL, MgCl2, PEG8000, Tween 20 and dithiothreitol; the joint combination comprises a joint sequence group and a tag sequence group. According to the kit, the library construction cost can be reduced and the amplification efficiency is improved.

Description

technical field [0001] The invention relates to the field of high-throughput sequencing library construction, in particular to a kit and method for constructing a DNA library. Background technique [0002] Now the common small fragment DNA library construction method mainly adopts NEB library construction kit, such as Ultra TM Or DNALibraryPrepKitforIllumina, etc. These kits are basically suitable for the construction of all small fragment (180bp ~ 500bp) libraries, including samples of different species, different quality or different fragment types. And the constructed library is widely used in de novo sequencing (denovosequencing) or resequencing (resequencing) and so on. [0003] The main process of library construction with the above kit is as follows: S1: Fragment the genome; S2: The end of the fragmented DNA is not blunt, so fill it in (20°C for 30 min), and then add A (65 ℃30min); S3: use TA connection to add a fixed sequence, that is, U-shaped linker (20℃15min),...

Claims

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Application Information

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IPC IPC(8): C40B50/06
CPCC40B50/06
Inventor 宗莹李宗文刘运超朱海浩王大伟李明洲宋莉
Owner BEIJING NOVOGENE TECH CO LTD
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