摘要
以椰子叶片为实验材料,探索了椰子基因组DNA的提取方法,并对影响RAPD反应的各因素进行了优化,建立了椰子的优化反应体系和程序。试验最终建立的椰子RAPD反应总体积为20μL,各有关成分的最佳浓度分别为Mg2+2.5 mmol/L,Taq DNA0.75 U,dNTP 0.2 mmol/L,Primer 1μmol/μL,模板DNA2.5 ng/μL。PCR循环程序为:94℃预变性1.5min;94℃变性20 s,36℃退火20 s,72℃延伸45 s,35个循环;最后72℃延伸3 min。
Using the leaves of coconut （ Cocos nucifera L. ） as the tested materials, the method of extracting genomic DNA was studied, and an improved technique was applied to extract the genomic DNA of coconut. The conditions for RAPD analysis were optimized, and the best RAPD reaction system was established as follows： total volume being 20 μL, containing 2.5 mmol/L Mg^2＋ , 0.75U Taq DNA polymerase, 0.2 mmol/L dNTPs, 1 μmol/μL primer, 2.5 ng/μL template DNA. PCR program was 1.5 min at 94 ℃ for predenaturalization, then 35 cycles of 20 s at 94 ℃ for denaturation, 20 s at 36 ℃ for annealing and 45 s at 72 ℃ for extension, finally extension at 72 ℃ for 3 min.
出处
《江西农业学报》
CAS
2008年第10期7-9,共3页
Acta Agriculturae Jiangxi
基金
国家科技基础条件平台重点项目"热带作物种质资源标准化整理
整合及共享试点"子项目"椰子种质资源标准化整理
整合及共享试点"(合同号:2005DKA21000-5-13)
中国热带农业科学院科技基金"香水椰子的苗期鉴定技术研究"(合同号:Rky0631)
