摘要
以红锥(Castanopsis hystrix A.DC.)嫩叶为材料,应用改良的CTAB方法成功提取了红锥基因组DNA,并对影响随机扩增多态DNA(RAPD)反应的各因素进行了优化,建立了红锥RAPD的优化反应体系及程序。在25μl反应体系中,模板DNA0.8ngμl-1,10×Buffer2.5μl,Mg2+2.0mmol/L,Taq DNA聚合酶0.8U,dNTPs0.35mmol/L,随机引物S420.28μmol/L。PCR循环程序为:94℃预变性3min,然后94℃30s,39℃1min,72℃2min,35个循环,最后72℃延伸10min,4℃保存。
The genomic DNA was extracted from Castanopsis hystrix A. DC. using the modified method of CTAB, and the optimization of its RAPD reaction system was also studied. Results showed that the high-quality genomic DNA can be obtained with the modified method of CTAB, and it could be directly used for RAPD analyses. The optimal PCR system for RAPD analysis was as follows： 0.8 ng μl^-1 DNA template, 2.0 mmol/L Mg^2＋, 0.8 U Taq polymerase, 0.35 mmol/L dNTPs, 0.28 μmol/L random primer S42, in 25 μl reaction system. RAPD program was 3 minutes at 94℃ for predenaturation, then followed by 35 cycles, each with 30 seconds at 94℃ for denaturation, 1 minute at 39℃ for annealing, 2 minutes at 72℃ for extension, finally extension at 72℃ for 10 minutes.
出处
《热带亚热带植物学报》
CAS
CSCD
北大核心
2008年第1期89-94,共6页
Journal of Tropical and Subtropical Botany
基金
广东省林业局2006年优良乡土树种选育和繁育专项基金资助
关键词
红锥
DNA提取
RAPD
体系优化
Castanopsis hystrix
DNA extraction
RAPD
System optimization
