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芝麻高度不育材料基因组DNA提取及RAPD反应体系的建立


芝麻高度不育材料基因组 DNA 提取及 RAPD 反应体系的建立 摘要 以临时保持系 WB51220 、两用系 0176A 不育株、可育株 0176B 和皖 芝 1 号等 4 份材料的叶片为研究材料, 对其基因组 DNA 的提取及对 RAPD 反应 体系进行优化。结果表明,改良的 CTAB 法获得的芝麻基因组 DNA 片段大小经 电泳检测满足 RAPD 等遗传多样性分析要求; 筛选出 RAPD 最佳反应体系: 20 μL 反应体系含有 1.5 U Taq DNA 聚合酶,0.25 mmoL/L dNTP,2.0 mmoL/L Mg2+, 0.75 μmoL/L 引物; 4 个不同品种的叶片基因组 DNA 的电泳条带之间有着明显差 异,他们共有的条带为 1 900、1 800、1 000、900 bp;不同的是,临时保持系 WB51220 比两用系 0176A 不育株少了 1 个 2 000 bp 的条带,可育株 0176B 比两 用系 0176A 不育株少了 1 个 450 bp 条带, 皖芝 1 号比可育株 0176B 少了 1 个 750 bp 和 1 个 1 200 bp 的条带。 Abstract The genomic DNA extraction and the RAPD-PCR reaction conditions were optimized when using genomes from sesame leaves of temporary maintainer line WB51220, AB Line infertile plant 0176A, fertile plant 0176B, and Wanzhi No.1 as templates. The results indicated that the genomic DNAs extracted from sesame leaves by improved CTAB DNA extraction method satisfied the requirement of genetic diversity analysis. The best reaction condition for RAPD analysis was as follows: Taq DNA polymerase 1.5 U, dNTP 0.25 mmoL/L, Mg2+ 2.0 mmoL/L, and random primer 0.75 μmoL/L in a total volume of 20 μL. Electrophoresis results indicated that bands amplified from the genomic DNAs of four varieties had four common bands in length of 1900,1800,1000,and 900 bp,respectively. The differences were that new bands in length of 2000 bp and 450 bp were amplified from AB line infertile plant 0176A when compared with temporary maintainer line WB51220 and fertile plant 0176B,respectively. And two new bands in le

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