Reagent for transposase fragmentation and application of reagent

Abstract

The invention discloses a reagent for transposase fragmentation. The reagent comprises the following assemblies independently packaged: Tn5transposase, a first connector, a second connector and a transposition reaction buffer solution. Further, the reagent also comprises a forward primer set, a reverse primer set, DNA polymerase, a DNA polymerase reaction buffer solution, as well as assemblies ofa fragmentation reaction buffer solution, a stop solution and the like, wherein the forward primer set, the reverse primer set, the DNA polymerase and the DNA polymerase reaction buffer solution are independently packaged. The invention also provides a reagent kit including the reagent. The reagent and the reagent kit provided by the invention are convenient to use, low in cost, good in use flexibility and suitable for various actual requirements, and the defects that a conventional commercialization transposition system is poor in stability, single in purposes, high in cost and the like can be avoided; and particularly when the reagent and the reagent kit are used for constructing a fragmentation nucleic acid library, the process of DNA fragmentation, terminal repair and connector connection can be completed only within 10min, the library construction time can be notably shortened, and excellent sequencing quality can be obtained.

Description

technical field [0001] The invention relates to a kit, in particular to a reagent and kit for transposase fragmentation, and belongs to the technical field of molecular biology equipment. Background technique [0002] The traditional library construction process is cumbersome, and generally involves the following steps: (1) Break up the DNA molecules to be sequenced with ultrasonic waves; (2) Fill in the ends of the broken genome; (3) Fill in the Phosphorylate the 3' end of the fragment by adding A and 5' end; (4) perform adapter ligation on the repaired fragment; (5) perform PCR amplification on the DNA fragment after adapter ligation; (6) perform PCR amplification on the amplified library Magnetic bead purification. [0003] Compared with the traditional library construction, the transposition method is simple and fast in its experimental method. The process of DNA fragmentation, end repair and adapter ligation is completed in one system, and the DNA fragmentation and ada...

AI Technical Summary

Problems solved by technology

However, the Tn5 transposition method library construction kits currently available on the market are generally only suitable for library construction, with a single scope of application, relatively high procurement costs, and long scheduled cycles.

[0004] For example, the commercialized transposition system provided by Illumina includes a mix system consisting of a transposase and two equimolar adapters (Adapter)1 and 2, which is expensive and designed for Illumina high-throughput sequencing Developed for a platform with a single purpose

[0005] In addition, existing commercial kits directly provide transposomes, which are less stable, and severe vibration and bumps during transportation may cause adapters to fall off, resulting in a decrease in fragmentation and affecting the quality and yield of the final library.

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