新疆野苹果基因组提取优化及模板浓度对ISSR-PCR影响  

Optimization of Genome Extraction and Effect of Template Concentration on ISSR-PCR in Malus sieversii

作　　者：张云秀[1] 杨美玲[2] 于玮玮[1] 李芳[1] 刘彬[1] 龙鸿[1] 阎国荣[1] 

机构地区：[1]天津农学院园艺园林学院,天津300384 [2]南开大学生命科学学院,天津300071

出　　处：《天津农学院学报》2017年第1期15-18,共4页Journal of Tianjin Agricultural University

基　　金：新疆维吾尔自治区财政林业科技专项项目"新疆野苹果种质资源收集与评价"(无编号)

摘　　要：以新疆野苹果干燥叶片为材料,比较分析了CTAB法和4种改良CTAB法提取基因组DNA效果,及ISSR-PCR扩增所需最佳模板浓度。结果表明:5种方法提取的DNA在纯度和量上有很大差别,研磨时加入PVP,可有效去除多酚物质;第一次提取上清液中加入1/10体积CTAB裂解液(0.7 mol/L Na Cl、10%CTAB),可以有效去除多糖;异丙醇沉淀时加入1/9体积的乙酸钠,可加速得到白色沉淀。新疆野苹果ISSR扩增体系研究中,发现DNA模板浓度处于400~1 000 ng/μL时,ISSR-PCR扩增结果最佳。DNA提取优化及模板浓度优化为后续的分子生物学研究提供理论依据。As Malus sieversii dried leaves for materials, compared and analyzed the result of extracting genomic DNA with the CTAB method and four kinds of modified CTAB method, besides, to determine the best template concentration for ISSR- PCR. Results showed that the purity and quantity of DNA with five methods has much difference, grinding with PVP can effectively remove polyphenols; addingl/10 volume CTAB cracking in the supernatant fuid （0.7 mol/L NaCl, 10% CTAB） can effectively remove polysaccharide; isopropyl alcohol precipitating with 1/9 volume sodium acetate can get white precipitation quickly. As for the M. sieversii ISSR amplification system study, found that ISSR-PCR amplified result is best with DNA template concentration in 400 - 1 000 ng/μL. DNA extraction and the concentration of template optimization provide theoretical basis for subsequent molecular biology research.

关 键 词：新疆野苹果 DNA提取 模板浓度 优化 

分 类 号：Q946-33[生物学—植物学]