安徽农业科学2016，Vol.44Issue(33)：137-139,142,4.
山楂PCR－SSCP反应体系与反应条件的优化
Optimization of PCR-SSCP Reaction System and Reaction Condition in Crataegus spp.
撒云俐 1那冬晨1
作者信息
- 1. 山西师范大学生命科学学院，山西临汾041000
- 折叠
摘要
Abstract
Objective] The aim was to optimize PCR-SSCP reaction system and reaction condition.[ Method] The chloroplast DNA and nu-clear DNA in Crataegus spp.were step-by-step extracted by improved CTAB method, the PCR-SSCP primers were selected, and the PCR-SS-CP reaction system and reaction conditions were optimizated.The PCR-SSCP productions were tested by agarose gelelectrophoresis, the dena-tured PCR-SSCP products were analyzed by native polyacrylamide gel.[ Result] The results showed that five pairs primers ( psbA-trnH、ropB、ropL、rpoC1、ITS2) of all are suitale for PCR-SSCP in Crataegus spp..The clear electrophoretogram of PCR-SSCP in Crataegus spp.were ob-tained through using 6% native polyacrylamide gel.(Crosslinking degree is 29∶1) at 4 ℃and 200 V for 3-4 h, with the volume of the load-ing buffer ( without glycerol) was same with the volume of PCR products, denaturation temperature was 98 ℃ under 1%NaOH, denaturetion time was 15 minutes, electrophoresis buffer was 0.5 ×TBE.[ Conclusion] The study optimize PCR-SSCP reaction system and reaction condi-tion, in order to lay the foundation for optimization of hawthorn SSCP analysis.关键词
山楂/PCR-SSCP/引物/反应体系/反应条件Key words
Crataegus spp/PCR-SSCP/Primers/Reaction system/Reaction conditions分类
农业科技引用本文复制引用
撒云俐,那冬晨..山楂PCR－SSCP反应体系与反应条件的优化[J].安徽农业科学,2016,44(33):137-139,142,4.
