Mutant Tn5 transposase and kit

Abstract

The invention provides a mutant Tn5 transposase and a kit, and belongs to the technical field of biology. On the basis of a wild type Tn5 transposase, the wild type Tn5 transposase has five amino acid site mutations: R26W, E54K, R62P, D156K and L372P, and at least two of three mutation sites: S19R, N204K and S267T, and the amino acid sequence of the wild type Tn5 transposase is as shown in SEQ ID NO: 1. The invention also relates to an application of the mutant Tn5 transposase in library construction. Compared with a wild type Tn5 enzyme, the mutant Tn5 transposase provided by the invention has the advantages that the activity is obviously improved, the preference on a DNA sequence is obviously reduced, dsDNA and DNA / RNA hybrid chains can be recognized without preference, any (0.1-50ng) input amount can be used for building a library within about 2.5 hours, and the requirements of various high-throughput sequencing library building can be met.

Description

technical field

[0001] The invention relates to a mutant Tn5 transposase and a kit, belonging to the field of biotechnology. Background technique

[0002] With the development of life science, we need to analyze the genetic information of multiple species. NGS, also known as high-throughput sequencing, can read the sequences of hundreds of thousands to millions of DNA molecules at one time, and can provide rich genetic information. The first step in NGS is library preparation, which is critical to the NGS workflow. This step produces a DNA or RNA sample that is compatible with the sequencer. DNA is usually fragmented first, and then specific adapters are added to both ends to construct a sequencing library.

[0003] At present, the methods of DNA fragmentation are mainly divided into physical fragmentation or enzymatic fragmentation. The enzyme cleavage method is increasingly becoming the preferred method for NGS library construction because of its high efficiency, conven...

Images