茶树ISSR-PCR反应体系优化研究
Optimization of ISSR-PCR reaction system in tea plant
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摘要: 通过优化影响茶树ISSR-PCR的主要参数,确立适合于茶树的ISSR反应体系和扩增条件。结果表明,在20μl反应体系中模板DNA的用量、Taq酶用量、引物浓度、Mg2+浓度、dNT p浓度分别为:模板DNA为40 ng;Taq酶为0.5 U;引物为0.25μmol·L-1;MgCl2为1.5 mmol·L-1;dNTp为0.2 mmol·L-1。适宜退火温度比Tm值高2~3℃。Abstract: The optimal ISSR-PCR reaction conditions in tea was established by studying the main parameters.Results showed that the optimal concentration of DNA,Taq polymerase,primer,Mg2+,dNTp in 20 μl reaction mixture was 40 ng,0.5 U,0.25 μmol·L-1,1.5 mmol·L-1,0.2 mmol·L-1.The appropriate annealing temperature was 2-3℃ higher than Tm of corresponding primer.
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