摘要
以南方菟丝子基因组DNA为试材,采用单因素递进筛选方法对菟丝子的RAPD反应体系进行了优化。结果表明,在25μL总反应体积中,最佳浓度配比为:模板DNA 20 ng,MgC l23.0 mmoL/L,引物10μmoL/L,dNTP 400μmoL/L,TaqDNA聚合酶1个单位(U),10×反应缓冲液2.5μL。扩增程序为:94℃变性3 m in,再进入循环;94℃变性1 m in,40℃退火1m in,72℃延伸1 m in,共40个循环;72℃最终延伸10 m in,于4℃保存。
The genomic DNA of Cuscuta australis was extracted and was used to optimize the RAPD reaction system of Cuscuta. The results indicated that ： the best reaction system was that 25 μL total reaction volmne of RAPD - PCR contained 20 ng template DNA, 3.0 mmol/L MgCl2, 10 μmoL/L primer,400 μmoL/L dNTP, 1 unit Taq DNA polymerase, 10 times buffer 2.5 μL. The thermal cycle condi, tion was： firstly denaturing at 94 ℃ for 3 minutes;then 40 cycles which involved denaturing at 94 ℃ for 1 minute,annealing at 40 ℃ for 1 minute, and elongation at72 ℃ for 1 minute;finally elongation at 72 ℃ for 10 minutes, and then was kept at 4 ℃.
出处
《江西农业学报》
CAS
2008年第5期21-24,共4页
Acta Agriculturae Jiangxi
基金
福建科技重大专项专题(2006NZ0002-1)
国家"十一五"重大科技支撑计划(2006BAD08A13)
