Rapid construction method of genome DNA sequencing library, and matched kit

Abstract

The invention discloses a rapid construction method of a genome DNA sequencing library, and a matched kit. The method comprises the following steps: (1) selecting DNase I deoxyribonuclease I as a DNAfragmenting enzyme to fragment a sample DNA by enzyme digestion to obtain fragmented DNA; (2) directly adding a dA tail to the fragmented DNA to obtain DNA with the dA tail; (3) connecting the DNA with the dA tail with a joint to obtain a joint connected product; (4) carrying out magnetic-bead purification on the joint connected product; (5) carrying out polymerase chain reaction (PCR) amplification on the joint connected product to obtain an amplification product; (6) carrying out magnetic-bead purification on the amplification product to obtain a genome DNA sequencing library product; and (7) carrying out fragment distribution inspection and library evaluation on the library product by adopting an agarose gel electrophoresis detection method. According to the invention, the DNase I is selected as the DNA fragmenting enzyme, so that the steps of tail end repair and phosphorylation are reduced, the library construction time is shorter, and cost is lower.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for rapidly constructing a genome DNA sequencing library and a matching kit. Background technique [0002] The acquisition of genetic information of living organisms is of great significance to the research of life sciences and promotes the rapid development of the field of biomedicine. In recent years, more and more genomes including human, Arabidopsis and rice have been sequenced, and whole genome sequencing has become an important means of life medical research. Research or clinical testing in various biological science fields involves the construction of sequencing libraries and high-throughput sequencing services. [0003] Conventional sequencing library preparation involves various steps such as fragmentation of sample DNA, end filling, phosphorylation at the 5' end, dA tailing at the 3' end, adapter ligation, and PCR amplification, and multiple steps of nucleic acid P...

AI Technical Summary

Problems solved by technology

However, for some staff who need to process a large number of database construction samples every day and who are new to database construction, the traditional database construction method is still too cumbersome

In particular, the fragmentation of DNA generally needs to be completed using a special ultrasonic fragmentation instrument. The cost of consumables is relatively high, and it cannot meet the needs of all users.

[0004] All in all, the existing methods for constructing genomic DNA sequencing libraries have the following defects: 1. Strong dependence on equipment and high cost; 2. Complicated operations, long time-consuming, and low efficiency of library construction; 3. Unable to achieve integrated operations; 4. Single sample Fragmentation, unable to achieve high-throughput library construction

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