黑果枸杞MSAP技术体系的构建  

Construction of MSAP technology system for Lycium ruthenicum Murr.

作　　者：刘晓雯 张得芳[1] 王占林[1] LIU Xiaowen;ZHANG Defang;WANG Zhanlin(Academy of Agriculture and Forestry Sciences,Qinghai University,Key Laboratory of Forest Genetics and Breeding of Qinghai Plateau,Xining 810016,China)

机构地区：[1]青海大学农林科学院,青海省高原林木遗传育种重点实验室,青海西宁810016

出　　处：《青海大学学报》2024年第2期35-42,共8页Journal of Qinghai University

基　　金：国家自然科学基金项目(32170239);青海省科学技术厅项目(2022-ZJ-725)。

摘　　要：为利用甲基化敏感多态性扩增技术(MSAP)构建黑果枸杞MSAP技术反应体系,本试验采用CTAB法提取黑果枸杞DNA,通过双酶切、预扩增和选择性扩增对16对引物的多态性进行初步筛选并构建MSAP技术体系。结果表明:双酶切进行12 h,DNA模板能够酶切完全;预扩增反应产物条带集中在250~1000 bp处,选择性扩增可筛选出6对引物组合,条带明显。本研究为后续黑果枸杞DNA甲基化水平的相关分析研究奠定了基础。Lycium ruthenicum Murr.DNA is extracted by adopting CTAB method in order to con-struct its methylation-sensitive amplified polymorphism(MSAP)technology reaction system utili-zing MSAP technology.Preliminary screening is conducted on the polymorphism of 16 pairs of pri mers by using double digestion,pre-amplification and selective amplification to construct MSAP technology system.The results show that DNA template can be digested completely by double diges-tion at 12 h.The bands of pre-amplification reaction products are concentrated at 250~1000 bp.In addition,six pairs of primer combinations are screened by selective amplification with obvious bands.This study lays a foundation for the further analysis of DNA methylation level of Lycium ru-thenicum Murr.

关 键 词：黑果枸杞 甲基化敏感多态性扩增技术(MSAP) DNA甲基化 引物组合 

分 类 号：Q691[生物学—生物物理学]