Optimization of RAPD reaction system of Taxus cuspidate Sieb. et Zucc | 东北红豆杉RAPD反应体系的优化
2010
Xiao Lu, Agricultural College of Yanbian University, Longjing（China）, Department of Agriculture | Xu Mingzi, Agricultural College of Yanbian University, Longjing（China）, Department of Agriculture | Cui Miao, Agricultural College of Yanbian University, Longjing（China）, Department of Agriculture
[Objective] The research aimed to study the optimization conditions of RAPD reaction system of Taxus cuspidata. [Method] Using the leaves of Taxus cuspidata as materials, DNA was extracted by using modified CTAB method. The influences of template DNA concentration, primer concentration, dNTP concentration, Taq DNA polymerase amount and magnesium ion concentration on the amplification effects of RAPD of Taxus cuspidate were analyzed. [Result] Through analysis and comparison of various factors, the optimized reaction system was established as follows: in a total volume of 20 μl, containing 10 ng DNA, 2.0 mmol/L Mg2+, 0.2 mmol/L dNTPs, 15 pmol/μl random primer, 1.0 U Taq polymerase, 2 μl 10×buffer, and the supplementary ddH2O. [Conclusion] The research provided technical support for further discussion on the related studies with the sexual identification of Taxus cuspidate.
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