河北工业科技2017，Vol.34Issue(1)：7-11,5.DOI:10.7535/hbgykj.2017yx01002
甘草SRAP-PCR正交试验设计优化及引物筛选
Optimization of SRAP-PCR system on Glycyrrhiza uralensis by orthogonal test design and selection of primers
摘要
Abstract
In order to establish the SRAP-PCR technology for Glycyrrhiza uralensis,an orthogonal design testing is used to optimize the reaction system of SRAP-PCR with 4 factors (Taq polymerase,Mg2+,dNTP and primer) at 4 levels,and the electrophoresis profiles are analyzed by software SPSS.The annealing temperature and SRAP primer combinations are selected using the single factor experiment.The results show that effects of the 4 factors on the SRAP-PCR are dNTP,Taq polymerase,Mg2+ and primer in turn.The optimized SRAP-PCR reaction system (20 μL) for G.uralensis is constructed of 1×PCR buffer,0.6 μmol/L primers,2.5 mmol/L Mg2+,1.5 U Taq polymerase,0.3 mmol/L dNTP and 40 ng DNA template.The optimized annealing temperature is 50 ℃.Using the optimized system,22 primer combinations are selected among 81 primer combinations,which produced bright and clear bands.The optimized SRAP-PCR system can provide the technique support for the phylogenetic analysis of G.uralensis.关键词
植物遗传学/甘草/SRAP/PCR/正交试验设计/引物筛选Key words
plant genetics/Glycyrrhiza uralensis Fisch./SRAP/PCR/orthogonal test design/primer screening分类
生物科学引用本文复制引用
艾鹏飞,苏姗,靳占忠..甘草SRAP-PCR正交试验设计优化及引物筛选[J].河北工业科技,2017,34(1):7-11,5.基金项目
河北省科技计划项目(14237503D-3) （14237503D-3）
