黄秋葵SRAP反应体系的优化

Optimizing SRAP Reaction System for Abelmoschus esculentus Linn.

摘要: 通过研究黄秋葵SRAP反应体系中主要因子TaqDNA聚合酶浓度、dNTP浓度、引物浓度和DNA模板浓度对扩增结果的影响,建立黄秋葵SRAP-PCR反应的优化体系。根据试验确定黄秋葵SRAP反应体系为20μL,75ng模板DNA,0.4μmol·L-1引物,0.750 mmol·L-1 dNTP,1.5 U Taq DNA聚合酶,2.0mmol·L-1 MgCl2,2.0μL 10×PCR缓冲液。

Abstract: Through optimization of the concentrations of Taq DNA polymerase,dNTP,primers and DNA template,SRAP-PCR of Abelmoschus esculentus Linn.was established.The results indicated that,in a 20μL reaction mixture,the optimal concentration of genomic DNA template was 75ng;that of the primers,0.4mmol·L-1;that of dNTP,0.75mmol·L-1;that of Taq DNA,polymerase,1.5U;and that of MgCl2,2.0mmol·L-1.