马尾松胚乳DNA高效提取及SSR-PCR体系优化  

Efficient DNA extraction from endosperm of Pinus massoniana and SSR-PCR system optimization

作　　者：何卫龙[1] 杨立恒[1] 王晓锋[1] 林能庆 季孔庶[1] 

机构地区：[1]南京林业大学林木遗传与生物技术省部共建教育部重点实验室,南京210037 [2]福建省上杭白砂国有林场

出　　处：《林业科技开发》2013年第1期15-18,共4页China Forestry Science and Technology

基　　金：林业公益性行业科研专项经费资助(编号:201104010);国家"十一五"科技支撑项目(编号:2006BAD01A1403)

摘　　要：以马尾松胚乳为试验材料,用CTAB-SDS法提取单粒胚乳DNA,对其浓度和纯度进行检测。结果表明:运用CTAB-SDS法提取的马尾松胚乳DNA浓度高、纯度好,琼脂糖凝胶电泳条带清晰,点样孔干净。此外,还对影响PCR结果的Mg2+、dNTP、模板DNA、引物浓度、TaqDNA聚合酶5个因素进行了正交设计试验,建立了适合马尾松SSR-PCR的反应体系:即在10μL反应体系中,Mg2+浓度为2.80 mmol/L,dNTP浓度为0.35 mmol/L,引物浓度为0.075μmol/L,模板DNA浓度为20 ng/10μL,TaqDNA聚合酶浓度为0.50 U/10μL。Take endosperm of P.massoniana as an experimental material and extract one endosperm DNA by CTAB-SDS,then,detect its concentration and purity.The experimental result indicated that the concentration and purity of DNA extracted from P.massoniana endosperm by CTAB-SDS were good,the agarose gel electrophoresis strips were clear and sample holes were clean.Besides,the experiment also took an orthogonal design towards Mg2＋,dNTP,template DNA,primer concentration,TaqDNA polymerase which could affect the PCR result.A suitable reaction system for P.massoniana SSR-PCR was established： in the 10 μl reaction system,2.80 mmol/L Mg2＋,0.35 mmol/L dNTP,0.075 mol/L primer,20 ng template DNA,0.50U TaqDNA polymerase.

关 键 词：马尾松 胚乳DNA CTAB-SDS法 SSR-PCR 

分 类 号：S661.2[农业科学—果树学]