鸭梨ISSR-PCR反应体系的初步建立

Establishment of ISSR-PCR reaction system of Pyrus bretschneideri Rehd. cv. Yali

以沧州泊头市鸭梨为试验材料,对鸭梨ISSR-PCR反应体系中的模板DNA用量、dNTPs浓度、引物浓度以及Taq酶浓度进行了探索,初步建立适合鸭梨ISSR-PCR反应体系为:在20μL反应体系中,含1×Taq PCR Buffer[1.5 mmol/L Mg2+]0、.15 mmol/L dNTPs、0.40μmol/L引物、40 ng DNA和1 U Taq酶。

The main influential factors of ISSR-PCR reaction system in Yali pear of Botou were researched.The effect of different concentrations of template DNA,primer,dNTPs and Taq DNA polymerase was analyzed.The results showed that the most suitable ISSR-PCR reaction system was 1×Taq PCR Buffer 1.5 mmol/L Mg2＋,0.15 mmol/L dNTPs,0.4 μmol/L primer 40ng DNA template,1.0 U Taq DNA polymerase in 20 μL reaction system.