南方红豆杉叶绿体非编码序列PCR体系优化及引物筛选  

Optimization of PCR Reaction System and Selection of cp DNA Noncoding Primers for Taxus chinensis var. mairei

作　　者：邹帆[1] 徐刚标[1] 何长青[1] 覃建庸[1] 

机构地区：[1]中南林业科技大学生命科学与技术学院,湖南长沙410004

出　　处：《广西林业科学》2016年第1期19-23,共5页Guangxi Forestry Science

基　　金：国家林业公益性行业科研专项(201104033)

摘　　要：PCR技术广泛用于分子标记、基因工程等领域,是植物系统发育及种群遗传结构等研究的基础。通过对影响PCR体系扩增的主要因子进行单因素试验,得出南方红豆杉(Taxus chinensis var.mairei)叶绿体DNA(cpDNA)最优体系(30μL)为3μL 10×PCR buffer,1μLDNA模板(60 ng),4.2μL MgCl_2(3.5 m M),8.4μLdNTPs(0.7 m M),0.9μL引物(0.3μM),0.8μL Taq DNA聚合酶(2 U),10.8μL去离子水。利用该体系筛选出叶绿体DNA非编码区通用引物4对,分别为atp I-atp H、psb J-pet A、trn H-psb A和trn L-trn F。PCR was used widely in fields of molecular markers and gene engineering, which was the basis of research on plant phylogeny and genetic structure of populations. The optimal cpDNA- PCR reaction system of Taxus chinensis var. mairei was established containing 3 μL 10 × PCR buffer, 1 μL template DNA （60 ng）, 4. 2 μL MgC12 （ 3.5 mM）, 8.4 μL dNTPs （ 0. 7 mM）, 0. 9 μL primers （0. 3 μM）, 0. 8 μL Taq DNA polymerase （2 U） and 10. 8 μL ddH20 in a total volume of 30 μL reaction system by single factor test of main factors influencing PCR system amplification. Using this system, 4 pairs of cpDNA noncoding primers which were atpI- atpH, psbJ-petA, trnH-psbA and trnL -trnF were selected.

关 键 词：南方红豆杉 叶绿体DNA 体系优化 引物筛选 

分 类 号：S791.49[农业科学—林木遗传育种]