黑果枸杞MSAP技术体系的构建
Construction of MSAP technology system for Lycium ruthenicum Murr.
刘晓雯 1张得芳 1王占林1
作者信息
- 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
- 折叠
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250～1 000 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.
Abstract
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 250～1 000 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Key words
Lycium ruthenicum Murr./methylation-sensitive amplified polymorphism(MSAP)/DNA methylation/primer combination引用本文复制引用
基金项目
国家自然科学基金(32170239)
青海省科技厅项目(2022-ZJ-725)
出版年
2024
NETL
NSTL
万方数据