摘要
对药用植物刺山柑遗传多样性研究分析,建立了刺山柑ISSR-PCR稳定的反应体系,利用植物基因组试剂盒提取刺山柑药材DNA;根据此引物采用正交实验设计考察影响PCR扩增的4个主要因素,筛选出最佳反应条件:25μL ISSR-PCR的最佳浓度为10×buffer 2.5μL、引物0.2μmol/L、模板40 ng/μL、dNTPs 0.4 mmol/L、Taq/DNA聚合酶0.5 U,利用梯度筛选获得U 808引物最佳退火温度。为利用这一分子标记对药用植物刺山柑进行遗传多样性分析奠定了基础。
Genetic diversity of medicinal plants of Capparis spinosa was researched, and ISSR stable reactionsystem was established in this study. Plant genomic DNA kit were used for extracting the total DNA of Capparis spinosa;according to the primers the 4 main factors that can affected the PCR amplification will be studied by the orthogonal experimental design,the optimal ISSR reaction was carried out in a volume of 25/μL containing 2.5 μL 10 × buffer, primer 0.2/μmol/L,template DNA 40 ng/μL, dNTPs 0.4 retool/L, Taq DNA polymerase 0.5 U, obtaining the best annealing temperature of primer U 808 by temperature gradient filter; The ISSR-PCR experiments has given good foundation for further study of Capparis spinosa.
出处
《种子》
CSCD
北大核心
2013年第2期38-41,共4页
Seed
基金
新疆维吾尔自治区科技支撑计划项目(编号:200933123)
