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

ABSTRACT

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 (5(4)) 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 micromol x L(-1) primer, 0.20 mmol x L(-1) dNTP s, 1.5 - 2.0 mmol x 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.