Method for detecting 3'-5' exo-activity of specific base by nuclease and kit thereof

Abstract

The invention discloses a method for detecting 3'-5' exo-activity of a specific base by nuclease and a kit thereof. The method firstly detects the exo-activity of the specific base by enzyme; the method can detect any base, including dATP, dCTP, dGTP, dTTP, dUTP and degenerate base, and the like, and the application range is wide; the method can detect the 3'-5' exo-activity of most nucleases; theuse of fluorescence as a detection signal avoids radioactive contamination and has high sensitivity; nucleotide chains modified by fluorophores cannot be required, and the cost is low; the method canbe used to rapidly and extensively screen nucleases, and the application is flexible and the flux is high.

Description

technical field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a method and a kit for detecting the 3'-5' excision activity of a nuclease on a specific base. Background technique

[0002] Many nucleases have 3'-5' exo-cutting activity. Detection of nuclease 3'-5' exonuclease activity is of great significance for understanding the correction ability of DNA polymerase and the degradation rate of DNA exonuclease. Nucleases have different exo-cutting abilities to different bases (including various modified bases, degenerate bases, etc.), and also have different exo-cutting abilities to bases under correct pairing and different forms of wrong pairing. Studying the exoclease activity of nucleases on a specific base is of great significance to molecular biology research and drug development.

[0003] Most of the existing techniques for detecting 3'-5' exonuclease activity require radioactive labeling, gel electrophoresis, enzyme-linked im...

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