摘要
采用正交设计和响应曲面设计 (RSD)优化适合于芦苇 [Phragmitesaustralis(Cav.) Trin.ex steud]随机扩增多态 DNA (RAPD)的反应体系 ,结果确定芦苇 RAPD反应体系的最佳方案为 :在 PCR扩增程序 ,冷启动 ,94℃变性 1 min,36℃复性 90 s,72℃延伸 2 min,4 0个循环 ;72℃延伸 7min,4℃保存 ;2 5 μl反应体系包括模板DNA 6 0 ng、三磷酸脱氧核苷酸 (d NTP) 7.5 mmol、Taq酶 1 .5 u、引物 5 pm
The optimum reaction systems of RAPD were obtained by orthogonal design and response surface design(RSD) The RAPD reaction system of Phragmites australis were determined For a 25 μl reaction,the optional composition included 60ng of template DNA,7 5 mmol of dNTP,1 5 u of taq DNA,5 pmol of primer The program for efficient amplification included 40 cycles of denaturation at 94℃ for 1 min,annealing at 36℃ for 1 5 min,and extension at 72℃ for 2 min(7 min for the final cycle)
出处
《广西科学》
CAS
2004年第2期148-150,160,共4页
Guangxi Sciences
关键词
芦苇
RAPD
正交设计
响应曲面设计
条件优化
Phragmites australis,RAPD,orthogonal design,response surface design,RAPD optimalAbstract Genome DNA was extracted from the tissue of the white spot virus infected cultured shrimp Penaeus monodon by TN homogenizing,DS digesting,proteinase K dec
