水产养殖专业毕业论文不同方法对鳟鱼基因组DNA提取效果的影响.doc

目 录 题 目 …………………………………………………………………… 2 摘 要 …………………………………………………………………… 2 1 文献综述 ……………………………………………………………… 2 1.1甘肃省虹鳟鱼种质资源 ……………………………………… 2 1.2国内外虹鳟鱼分子水平研究进展 …………………………… 2 1.3研究目的及意义 …………………………………………………… 3 2 材料与方法 …………………………………………………………… 3 2.1实验材料 …………………………………………………………… 3 2.1.1实验样品 ………………………………………………………… 3 2.1.2主要仪器 ………………………………………………………… 3 2.1.2主要试剂 ……………………………………………………… 4 2.1.4溶液配制 ……………………………………………………… 4 2.2基因组DNA提取方法 ……………………………………………… 5 2.3基因组DNA检测 …………………………………………………… 6 2.3.1琼脂糖凝胶电泳检测 ………………………………………… 6 2.3.2 DNA纯度及浓度测定 ………………………………………… 6 3 结果与分析 ………………………………………………………… 6 3.1琼脂糖凝胶电泳检测结果 ……………………………………… 6 3.2 ND-1000型分光光度计检测结果………………………………… 7 4 讨论 …………………………………………………………………… 9 5 结果 ………………………………………………………………… 10 参考文献 ……………………………………………………………… 11 摘要………………………………………………………………… 13 致 谢 ………………………………………………………………… 14 不同方法对鳟鱼基因组DNA提取效果的影响 摘要 为虹鳟鱼遗传特征研究奠定基因组DNA提取的实验基础，本研究以虹鳟鱼肌肉组织为材料,按照常规的酚-氯仿抽提法和另外设计的一种不同方法进行基因组DNA提取并比较其效果。结果表明, 采用常规酚-氯仿抽提法提取的基因组DNA受蛋白质污染小；而另一种未加蛋白酶K的DNA提取法由于蛋白质未被消化降解且抽提次数较少，虽提取的DNA浓度较高但蛋白质残留较为严重，这对于PCR扩增的影响较大。因此，使用蛋白酶K的常规酚-氯仿抽提法提取的基因组DNA适于PCR扩增等后续实验研究。 关键词 鳟鱼；基因组DNA；提取；效果；比较 Effects of Different Methods on Extracting Genomic DNA in Trout Abstract: For genetic characteristics study lays didn genomic DNA extracted, this study experimental basis for materials, didnt muscle tissue according to conventional phenolic - chloroform extraction formulation and other design of a different methods for genomic DNA extracted and compares their effect. The results showed that the conventional phenolic - chloroform extraction of genomic DNA extracted by latifah protein pollution is small; And another didnt add proteinase K DNA extraction has not been digesting degradation due to protein and extraction, although fewer extracted DNA high concentration but protein relatively serious, the residual effects of for PCR bigger. Therefore, using proteinase K conventional phenolic - chloroform extraction of genomic DNA extracted for formulation of subsequent experiment research