摘要
为建立高效的筇竹SSR反应体系,以筇竹新鲜幼嫩叶片提取的基因组DNA为模板,研究了影响SSR反应效果的因素,包括Taq酶、Mg2+、dNTPs、引物、模板DNA浓度等,建立了适合筇竹SSR反应的PCR体系,即20μL反应体系中含有Taq酶0.8 U,Mg2+为1.5 mmol.L-1,dNTP 0.2 mmol.L-1,引物0.25μmol.L-1,模板DNA60 ng.扩增程序为:94℃3 min,然后94℃40 s,52℃30 s,72℃45 s,30个循环,72℃延伸5 min,4℃保存.
Factors that influence SSR reactions including Taq polymerase,Mg2＋,dNTPs,primers,template DNA＇s concentration and so on were studied using fresh young leaves of Qiongzhuea tumidnoda.An optimal PCR system for SSR of Qiongzhuea tumidnoda has been established：in 20 μL reaction solution,contained 0.8 U Taq-polymerase,1.5 mmol·L-1 of Mg2＋,0.2 mmol·L-1 of dNTPs,0.5 μmol·L-1 of primers,and 60 ng of template DNA.Amplification program was devised：initial denaturing at 94 ℃ for 3min,denaturing at 94 ℃ for 40 s,annealing at 52 ℃ for 30 s,and extension at 72 ℃ for 45 s,30 cycles;ending with a final extension at 72 ℃ for 5 min,then stored at 4 ℃.
出处
《河南农业大学学报》
CAS
CSCD
北大核心
2010年第6期644-648,共5页
Journal of Henan Agricultural University
基金
国家"十一五"林业科技支撑计划专题(2006BAD19B0202)
国家林业局重点项目(2006-64)
关键词
筇竹
SSR
体系优化
Qiongzhuea tumidnoda
SSR
Optimization of system
