摘要
以福鼎大白茶为材料 ,采用美国的PTC 10 0 Tm热循环仪 ,对茶树RAPD分析中影响PCR扩增结果的主要因素进行了研究 ,确定了茶树RAPD的最适反应体系和扩增程序 ,即在 30 μl反应体系中 ,含 40ng模板 ,2mmol/LMgCl2 ,各 0 .2mmol/LdNTPs、0 .15 μmol/L引物和 1.5UTaqDNA聚合酶 ;扩增程序为 :第 1步预变性94℃ ,180s ;第 2步变性 92℃ ,5 0s;第 3步退火 35℃ ,5 0s ;第 4步链延伸 72℃ ,10 0s ;循环 41次 ,后延伸 72℃ ,30 0s .
The optimal reaction mixture and amplification procedure of RAPD in Fuding Dabai Cha[Camellia sinensis(L.)O.Kuntze] was studied with American thermocycle machine(Model-PTC-100 Tm ). The results showed that each 30?μl amplification reaction solution was consisted of 40?μg template DNA,2?mmol/L MgCl 2,0.2?mmol/L dNTP of each,0.15?μmol/L primer and 1.5 U Taq DNA polymerase. The amplification procedure conditions were predenature at 94?℃ 180?s followed by denature at 92 ℃ 50?s, annealing 35?℃ for 50?s, extension 72?℃ for 100?s, cycling 41 times, last extension 300?s and then the optimum procedure for tea plant DNA amplification was obtained.
出处
《生命科学研究》
CAS
CSCD
2002年第2期179-182,共4页
Life Science Research
基金
湖南省自然科学基金资助项目 (OOJJY2 0 10 7)
