栝楼的RAPD-PCR体系建立与优化

目的]建立栝楼的RAPD-PCR体系并对该体系进行优化。方法]以栝楼叶片为材料,采用CTAB法提取栝楼叶片基因组DNA,利用正交设计对RAPD-PCR体系进行优化。结果]各因素水平变化对PCR反应影响大小依次为：Mg2＋、TaqDNA聚合酶、引物和dNTPs。通过试验分析,优化的栝楼RAPD反应体系为：在25μl反应体系中,含10×buffer2.5μl,Mg2＋2.0mmol/L,Taq酶1U,引物0.8μmol/L,dNTPs0.1mmol/L。反应程序为94℃预变性2mim;94℃变性30s,37℃退火温度40s,72℃延伸1.5mim,36次循环;72℃延伸10mim,4℃保存。结论]该研究建立的栝楼RAPD反应体系稳定可靠,为栝楼性别鉴定、遗传多样性分析、亲缘关系分析等方面的研究提供了有效的方法。

Establishment and Optimization of RAPD-PCR System in Trichosanthes kirilowii Maxim

Objective] The aim was to establish and optimize the RAPD-PCR system of Trichosanthes kirilowii Maxim. Method] With the leaves of T. kirilowii as materials, the genomic DNA of T. kirilowii leaves were extracted by CTAB method and the orthogonal design was used to optimize the RAPD-PCR system of T. kirilowii. Result] The effects of level changes of each factor on PCR reaction in the following order was Mg2+, Taq DNA polymerase, primer and dNTPs. Through experimental analysis, the optimized RAPD-PCR system...