黔产苦参ISSR-PCR反应体系正交优化研究
Orthogonal Optimization of ISSR-PCR Reaction System of Suphora flavercens Ait
金燕 1龙庆德 1常楚瑞 1陆平祝 1王晓丽1
作者信息
摘要
[目的]确定适合苦参稳定的ISSR-PCR反应体系。[方法]采用经典的CTAB法提取苦参新鲜幼嫩叶片DNA；针对Mg2+、dNTPs、引物、模板DNA和Taq DNA聚合酶进行正交设计L16(45)，优化黔产苦参ISSR-PCR反应体系。[结果]最适ISSR-PCR反应体系为：模板DNA 90 ng、0.4μmol/L引物、2.0 mmol/L Mg2+、Taq DNA聚合酶0.5 U、0.5 mmol/L dNTPs。[结论]建立的苦参ISSR-PCR反应体系，经过19份苦参样品检验，证明该体系稳定可靠，可用于苦参遗传多样性分析。
Abstract
Objective]To select a suitable method of ISSR-PCR reaction system for Suphora flavercens Ait. [Method] Modified CTAB method was used to extract the genomic DNA from fresh young leaves of S. flavercens. The suitable ISSR-PCR reaction system was established by orthogo-nal design L16(45) based on the Mg2+, dNTPs, primer,template DNA and Taq DNA. [Result]The optimal conditions in 25μL ISSR-PCR system were 90 ng DNA template, 0. 4μmol/L primer, 2. 0 mmol/L Mg2+, 0. 5 U TaqDNA Polymerase, and 0. 5 mmol/L dNTPs. [Conclusion]The es-tablished ISSR-PCR reaction system is verified by 19 samples of S. flavercens, which verifies that this system is stable and reliable, and can be used for the analysis of genetic diversity of S. flavercens.
关键词
苦参/CTAB/ISSR-PCR反应体系/正交设计Key words
Suphora flavercens Ait/CTAB/ISSR-PCR reaction system/Orthogonal test引用本文复制引用
基金项目
贵州省中药现代化科技产业研究开发专项(黔科合ZY字[2012]3004号)
贵州省科技计划项目(黔科合重大专项字[2015]6009-2)
出版年
2016
国家科技期刊平台
NSTL
维普
万方数据