陈山红心杉SCoT反应体系建立及引物筛选  

Establishment and optimization of SCoT-PCR reaction system and selection of primers for Red-heart Cunninghamia lanceolata

作　　者：娄永峰[1] 宋晓琛[1] 冷春晖 陈兴彬 朱新传 肖复明[1] Lou Yongfeng;Song Xiaochen;Leng Chunhui;Chen Xingbin;Zhu Xinchuan;Xiao Fuming(Jiangxi Provincial Key Laboratory of Plant Biotechnology,Jiangxi Academy of Forestry,Nanchang Jiangxi 330013,China;Chenshan Forest Farm of Anfu County,Anfu Jiangxi 343214,China)

机构地区：[1]江西省林业科学院·江西省植物生物技术重点实验室,江西南昌330013 [2]安福县陈山林场,江西安福343214

出　　处：《南方林业科学》2021年第4期1-4,32,共5页South China Forestry Science

基　　金：江西省林业科学院院博士启动项目(项目编号:2017521102);江西省林业局科技创新专项〔2021〕11号。

摘　　要：研究采用正交设计及单因素相结合的方法建立优化陈山红心杉SCoT-PCR反应体系,并在此基础上进行引物筛选。结果表明:SCoT-PCR反应的最优体系(20.0μL)为:模板DNA 2.5μL(50.0 ng),2×Hieff PCR Master Mix 9.0μL,引物3.0μL(即浓度1.5μmol·L^(-1));并在此基础上从36条SCoT引物中筛选出14条扩增条带丰富明亮且多态性较强的引物。这为今后利用SCoT分子标记研究陈山红心杉遗传多样性、种质资源鉴定等提供基础。The combination of orthogonal design and single factor experiment design were used to optimize the SCoT-PCR reaction system of Red-heart Cunninghamia lanceolata,and on this basis,the primers were screened.The results showed that the optimal reaction system of SCoT-PCR for Red-heart C.lanceolata was as follows:2.5μL(50.0 ng)template DNA,9.0μL 2×Hieff PCR Master Mix,3.0μL(1.5μmol·L^(-1))primers,and ddH2O supplemented to 20.0μL;14 primers with clear amplification bands and high polymorphism were screened out from 36 SCoT primers.It can provide relevant basis for the genetic diversity,identification of germplasm resources of Red-heart C.lanceolata.

关 键 词：SCoT 体系优化 正交设计 陈山红心杉 

分 类 号：S791.27[农业科学—林木遗传育种]