水稻TRAP-PCR反应体系优化与P-糖蛋白基因片段的分析

Optimization of TRAP-PCR Reaction System and Analysis of P-glycoprotein Gene in Rice

对影响TRAP-PCR反应体系的各参数,包括模板DNA、dNTP、TaqDNA聚合酶和引物浓度进行了优化,建立了适合水稻的稳定,可重复的TRAP-PCR反应体系。在20μLPCR反应体系中,含80ng模板DNA,0.25mmol/LdNTP,0.75UTaqDNA聚合酶,5ng/μL随机引物和7.5ng/μL特异引物。本研究对P-糖蛋白基因片段进行了克隆并序列分析,为深一步研究奠定了基础。

The factors that might influence TRAP-PCR, concentration of genomic DNA template, dNTP mixture, Taq DNA polymerase, primers, were optimized in this paper, and the advanced reaction system for TRAP-PCR in rice were established, that is total 20 μL reaction volume including 80 ng genomic DNA template, 0.25 mmol/L dNTP mixture, 0.75 U Taq DNA polymerase, 5 ng/μL arbitrary and 7.5 ng/μL specific primer. We also isolated a fragment of P-glycoprotein gene and then sequenced, which would be a basis for our further study.