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[Study on optimization of SRAP-PCR reaction system for Pinellia ternata in Suzhou]
Ai-Min Zhang1, He-Dong Lu, Jian-Ping Xue
1Anhui Key Laboratory of Plant Resources and Biology, School of Life Science, Huaibei Normal University, Huaibei 235000, China.
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.
