Establishment of ISSR marker technology and optimization of its system in Prunella vulgaris
- VernacularTitle:夏枯草ISSR分子标记技术的建立与体系优化
- Author:
Li LIAO
;
Qiaosheng GUO
- Publication Type:Journal Article
- Keywords:
Prunella vulgaris L.;
ISSR-PCR;
orthogonal design;
single factor test
- From:
Chinese Traditional and Herbal Drugs
1994;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish and optimize the ISSR-PCR reaction system for Prunella vulgaris and lay foundation for its genetic diversity research.Methods The single-factor and orthogonal design were applied for optimizing seven factors in the ISSR-PCR reaction system including Mg2+,dNTP,primers,Taq DNA polymerase,the template DNA,annealing temperature,and cycles.Results The suitable PCR reaction system contained 2.2 mmol/L Mg2+,175 ?mol/L dNTP,0.75 ?mol/L primer,1.0 U Taq DNA polymerase,and 30 ng template DNA in total 20 ?L reaction solution.On this basis,18 primers were screened with stable amplification and rich polymorphism from 92 ISSR primers.The optimal annealing temperature for ISSR-PCR reaction was proposed by gradient PCR.Conclusion It is a way to establisb the ISSR-PCR system for orthogonal design combining with single-factor test.And it is proved to be stable and credible for the result of 24 P.vulgaris populations.This optimized ISSR reaction system would provide the basis for the genetic analysis of P.vulgaris.
