[Study on optimization of SRAP-PCR reaction system for Pinellia ternata in Suzhou]

Objective: To investigate the optimization system of SRAP-PCR molecular marker technology in the analysis on Pinellia ternata.Method: SRAP-PCR reaction system for P.ternata was optimized by L16(54) orthogonal design with five elements(dNTPs,Mg2+,the template DNA,primers,Taq enzyme) and four standards.Result: The most suitable forward primer for SRAP for Pinellia ternata was 5′-TGAGTCCAAACCGGAAG-3′,while the reverse primer was 5′-GACTGCGTACGAATTACG-3′.The optimized reaction system contained 70 ng DNA template,0.9 μmol·L-1 primer,0.20 mmol·L-1 dNTP s,1.5~2.0 mmol·L-1 Mg2+,and 2 U Taq enzyme.Conclusion: SRAP-PCR system for P.ternata is established to lay a foundation for future construction of SRAP genetic map of P.ternata.