Optimization of SRAP-PCR reaction systems for Curcuma wenyujin / 中草药

ABSTRACT

Objective: To research on the most appropriate SRAP-PCR amplication system of Curcuma wenyujin. Methods: The concentration of Mg2+, dNTP, template DNA, primer, and Taq polymerase was optimized by amplifying technology and method of PCR based on isolating genomic DNA from C. wenyujin. Results: The best amplification system for C. wenyujin was as follows: total reaction volume of 25 μL, including Mg2+ (25 mmol/L) 2.0 μL, Taq polymerase(5 U/μL)0.4 μL, random primer (20 μmol/L) 2 μL, template DNA (5 ng/μL) 2.0 μL, dNTP (25 mmol/L) 2.0 μL, 10 × PCR buffer 2.5 μL. Conclusion: The stripes of amplification products are clearness, high brightness, and good reproducibility. It indicates that the reaction system and reaction procedures which are confirmed in this experiment are applied to the SRAP molecular markers in C. wenyujin, based for the genetic diversity of C. wenyujin.