Sequencing library construction method, and kit and application thereof

Abstract

The invention discloses a sequencing library construction method, and a kit and an application thereof. The method comprises the following steps: target DNA and a transposase-embedding complex are incubated under transposition reaction conditions, such that a DNA library with double linkers on the two ends is produced, wherein the transposase-embedding complex comprises transposase, a transposase recognition sequence complementary sequence, a first sequencing linker sequence and a second sequencing linker sequence. The first sequencing linker sequence comprises a first sequencing tag sequence at the 5' end and a transposase recognition sequence at the 3' end. The second sequencing linker sequence comprises a second sequencing tag sequence at the 5' end, a sample tag sequence and a transposase recognition sequence at the 3' end. With the method provided by the invention, with transposase, DNA can be broken with a one-step method, and sequencing linkers can be respectively added to the 5' end and the 3' end. Compared with a conventional library construction method, the method is time-saving, and the operation is simple.

Description

technical field [0001] The invention relates to the technical field of sequencing, in particular to a method for constructing a sequencing library, a kit and an application. Background technique [0002] With the advantages of high throughput and low cost, next-generation sequencing technology has been popular since its appearance. With the development of technology, next-generation sequencing technology has been applied in many scientific research and clinical testing. [0003] At present, many scientific researches and clinical applications need to be carried out at the single cell level, or at the trace level. It is also a common research method to analyze DNA genetic variation information at the single-cell level to determine whether a cell, embryo or individual is diseased or carries a disease gene. For example, preimplantation genetic diagnosis (Preimplantation Genetic Diagnosis) in assisted reproductive technology involves DNA genetic testing of gamete cells, single...

AI Technical Summary

Problems solved by technology

However, since the DNA fragmentation and adapter addition achieved by transposase can only be tagged at the 5' end of the target DNA, a step of adding a tag sequence at the 3' end catalyzed by a nucleic acid modifying enzyme such as DNA polymerase is required to obtain Double adapter DNA library, which inevitably increases the time-consuming of library construction

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