Kit and method for constructing DNA library

Abstract

The invention provides a kit and a method for constructing a DNA library, the kit comprises a mixed enzyme solution, a mixed buffer solution, ligase and a ligation buffer solution, the mixed enzyme solution comprises endonuclease, T4 DNA polymerase, Klenow fragments, Taq DNA polymerase and T4 polynucleotide kinase, the mixed buffer solution and the ligation buffer solution comprise DTT and PEG 8000, the mixed enzyme solution and the mixed buffer solution are used for forming a first reaction system, the ligase and the ligation buffer solution are used for forming a second reaction system, in the first reaction system, the working concentration of the endonuclease is 0.01-0.05 U / mu L, the working concentration of the T4DNA polymerase is 0.04-0.1 U / mu L, the working concentration of the Klenow fragment is 0.005-0.02 U / mu L, the working concentration of the Taq DNA polymerase is 0.025-0.075 U / mu L, the working concentration of the T4 polynucleotide kinase is 0.1-0.5 U / mu L, the working concentration of the DTT is 5-10mM, and the working concentration of the PEG 8000 is 1-10%. According to the invention, the kit and the method for constructing the DNA library, which can improve the library construction efficiency, can be provided.

Description

technical field [0001] The present disclosure particularly relates to a kit and method for constructing a DNA library. Background technique [0002] It is well known that next-generation sequencing (NGS) systems can perform a large number of parallel sequencing reactions simultaneously. Although different next-generation sequencing platforms have many differences in technical details, libraries need to be established before sequencing, so the acquisition of libraries is very important. [0003] At present, library construction kits (such as those provided by manufacturers such as illumina, Roche, NEB, etc.) are usually used to construct libraries, and the main process of library construction includes: fragmenting the genome, end repair, adding A reaction, ligation Adapter, magnetic bead capture fragment, PCR amplification and purification steps. However, these library construction kits often have problems such as complicated library construction process, long experiment ti...

AI Technical Summary

Problems solved by technology

However, these library construction kits often have problems such as complicated library construction process, long experiment time, and low library delivery rate.

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