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最新整理资料 文档精选合集 课件下载地址厦门大学公共卫生学院主页→学院概况→学科建设→病原微生物与 抗感染治疗课题组 \h 课件下载地址 厦门大学公共卫生学院主页→学院概况→学科建设→病原微生物与 抗感染治疗课题组 \h /s/81/t/263/08/c4/info133316.htm 网页下方的课件下载：MBPH1-10 Molecular Biology and Public Health （分子生物学与公共卫生） ------DNA/RNA manipulation 1 （DNA/RNA 操作1） IntroductionThe methods depend upon, and are developed from, an understanding of the properties of biological macromolecules Introduction The methods depend upon, and are developed from, an understanding of the properties of biological macromolecules themselves. 3 Nucleic acids 1.Isolation/Extraction（提取） 2.Electrophoresis（电泳） 3.Restriction（限制性酶切） 4.Hybridization（杂交） 5.PCR（聚合酶链式反应） 6.Genome sequence & analysis（基因 组测序&分析） 7.DNA recombination （DNA重组概念） 8.DNA recombination technology and cloning（ DNA重组和克隆技术) ReagentsPhenol ( Reagents Phenol ( 苯 酚 ): The phenol used for biochemistry comes as a water- saturated solution with Tris buffer, as a Tris-buffered 50% phenol, 50% chloroform solution, or as a Tris-buffered 50% phenol, 48% chloroform, 2% isoamyl alcohol solution (sometimes called "25:24:1"). Phenol is naturally somewhat water-soluble, and gives a fuzzy interface, which is sharpened by the presence of chloroform, and the isoamyl alcohol reduces foaming. Most solutions also have an antioxidant, as oxidized phenol damages the nucleic acids. For RNA purification, the pH is kept at around 4, which retains RNA in the aqueous phase preferentially. For DNA purification, the pH is usually near 7, at which point all nucleic acids are found in the aqueous phase. Chloroform（ 氯 仿 ）: Chloroform is stabilized with small quantities of amylene or ethanol, because exposure of pure chloroform to oxygen and ultraviolet light produces phosgene gas. Some chloroform solutions come as pre-made a 96% chloroform, 4% isoamyl alcohol mixture that can be mixed with an equal volume of phenol to obtain the 25:24:1 solution. Isoamyl alcohol（异戊醇 ）: Isoamyl alcohol may reduce foaming and ensure inactivation of RNases. How it works?Phase sep