Method and kit for detecting forest tree xylem DNA methylation through MSAP technology

Abstract

The invention discloses a method and a kit for detecting forest tree xylem DNA methylation through an MSAP technology. The method includes following steps: (S1): extracting forest tree xylem DNA; (S2) performing enzyme digestion to the forest tree xylem DNA with two enzyme sets: EcoRI / HpaII and EcoRI / MspI; (S3) connecting a linker of a restriction enzyme onto a product obtained through the enzyme digestion; (S4) performing PCR pre-amplification to a pre-amplification primer designed with the linker as a template to obtain a PCR pre-amplification product; (S5) diluting the PCR pre-amplification product and performing PCR amplification with addition of a screening primer having selective basic groups; and (S6) carrying out electrophoresis detection to a PCR amplification product and counting and analyzing DNA bands. By means of the technical scheme, a strong technical support is provided for researching of epigenetic variation so that a new candidate marking locus can be provided for targeted forest tree molecular marker assistant breeding.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a method and a kit for detecting DNA methylation of forest xylem by using MSAP technology. Background technique [0002] Epigenetic regulation is a molecular genetic mechanism that does not depend on genome sequence changes, but changes phenotypes through chromatin structure, base structure variation, and remodeling. It plays an important role in genome defense, gene expression, and nuclear inheritance. effect. Genomic DNA methylation is one of the most in-depth epigenetic mechanisms studied so far (Dahl and Guldberg, Biogerontology, 2003, 4: 233–450), and its detection methods are diverse and developing rapidly. For: methylation-sensitive endonuclease digestion, bisulfate treatment of methylated cytosine, polymerase chain reaction (PCR) and biochips, etc. [0003] Methylation-sensitive amplification polymorphism (MSAP) technology is a relatively mature, stable and re...

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Problems solved by technology

[0006] The present invention aims to provide a method and a kit for detecting DNA methylation of forest xylem by using MSAP technology, so as to solve the technical problem in the prior art that there is no method for stably and efficiently detecting DNA methylation of forest xylem

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