摘要
以中国荷斯坦奶牛为实验材料,研究了Mg2+浓度、dNTPs浓度、引物浓度、模板DNA浓度、TaqDNA聚合酶,以及退火温度对RAPD反应的影响,建立一套适合中国荷斯坦牛的最佳RAPD反应体系。反应总体积为20μL,Mg2+浓度为2.5mmol/L,TaqDNA聚合酶浓度为1U,引物浓度为2μmol/L,dNTPs浓度为400μmol/L,模板DNA浓度为100ng。PCR反应程序:94℃预变性2min→40循环(94℃变性1min→36℃退火1min→72℃延伸1min)→72℃延伸5min。
In this paper, the effects of concentration of Mg^2＋, dNTPs, random primers, template DNA,TaqDNA poiymerase and anneal temperatire on RAPD were analyzed with Holstein cattle. An optimalized RAPD system with total volume 20μL including Mg^2＋ at 2.5mmol/L, TaqDNA polymerase at 1 U, ramhom primers at 21℃/L, dNTP at 400μmol/L, template DNA at 100ng.The reaction pragram with 2. minutes at 94℃,which is followed by 40 cycles .each with 1 minutes at 94℃. ,1 minulcs al 36℃ ,1 minutes an 72℃ ,and a final extension at 72℃ for 5 minutes.
出处
《畜禽业》
2006年第3期8-11,共4页
Livestock and Poultry Industry
关键词
奶牛
RAPD
反应条件
Dairy Cattle
RAPD
Reaction Condition
