宿半夏SRAP PCR反应体系优化的研究(3)
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Study on optimization of SRAP-PCR reaction System for
Pinellia ternata in Suzhou
ZHANG Ai-min1， LU He-dong1， XUE Jian-ping1*， TAO Xing-kui1，2， XUE Tao1， SHENG Wei1， ZHU Yan-fang1
(1. Anhui Key Laboratory of Plant Resources and Biology， School of Life Science， Huaibei Normal University，Huaibei 235000， China； 2. Chemistry and Life Science Department， Suzhou University， Suzhou 234000， China)
[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(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.
[Key words] Pinellia ternata； SRAP-PCR； optimization of reaction system； orthogonal design
doi：10.4268/cjcmm20122428
[责任编辑 孔晶晶], 百拇医药(张爱民,卢河东,薛建平,陶兴魁,薛涛,盛玮,朱艳芳)
Study on optimization of SRAP-PCR reaction System for
Pinellia ternata in Suzhou
ZHANG Ai-min1， LU He-dong1， XUE Jian-ping1*， TAO Xing-kui1，2， XUE Tao1， SHENG Wei1， ZHU Yan-fang1
(1. Anhui Key Laboratory of Plant Resources and Biology， School of Life Science， Huaibei Normal University，Huaibei 235000， China； 2. Chemistry and Life Science Department， Suzhou University， Suzhou 234000， China)
[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(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.
[Key words] Pinellia ternata； SRAP-PCR； optimization of reaction system； orthogonal design
doi：10.4268/cjcmm20122428
[责任编辑 孔晶晶], 百拇医药(张爱民,卢河东,薛建平,陶兴魁,薛涛,盛玮,朱艳芳)