Optimization for RAPD-PCR reaction system of Bupleurum chinense DC. by comprehensive scoring method
- VernacularTitle:综合评分法优化柴胡RAPD-PCR反应体系
- Author:
Min YANG
;
Jiying HU
;
Keli CHEN
;
Yang BAI
- Publication Type:Journal Article
- Keywords:
RAPD;
Bupleurum chinense DC;
orthogonal design;
comprehensive scoring method
- From:
Chinese Traditional Patent Medicine
1992;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To optimize the random amplified polymorphic DNA-Polymerase chain reaction(RAPD-PCR) system of Bupleurum chinense DC by comprehensive scoring method. METHODS: The L_(16)(4~5) orthogonal diagram was applied to studying the effects of different concentrations of Taq polymerase,Mg~(2+),dNTPs,primer and DNA on Bupleurum chinense DC.RAPD-PCR results and to get a suitable PCR system. RESULTS: A suitable RAPD reaction system was developed,i.e.1?Taq polymerase reaction buffer,2.5 mmol/L Mg~(2+),1.5 U Taq DNA polymerase,0.15 mmol/L dNTPs,0.4 ?mol/L primer and 25 ng genomic DNA templates in total 25 ?L reaction solution. CONCLUSION: The results show that the comprehensive scoring method could be used successfully to optimize the RAPD-PCR system.
