提取和分离课件VIP

1、Figure3.1 The basic steps in preparation of total cell DNA from a culture of bacteriaBasic Steps基本步骤：1) 收集细菌细胞2)打碎细胞，释放内容物3)去掉DNA以外的成分4)DNA溶液的浓缩37C，150-250rpm达到最大浓度2-3109cell/ml, 600nm下的OD值，1OD相当于0.8109cell/ml Figure3.2 Estimation of bacterial cell number by measurement of optical densityFigure3.3 H

2、arvesting bacteria by centrifugationFigure3.4 Preparation of a cell extract.Figure3.5 Removal of protein contaminants by phenol extraction. Figure3.6 Collecting DNA by ethanol precipitation .Figure3.7 The CTAB method for purification of plant DNA十六烷基三十六烷基三甲基溴化铵甲基溴化铵Figure3.8 The use of an anion-exch

3、ange chromatography resin in DNA purification.Figure3.9 Preparation of a cleared .Sphaeroplast残壁细胞残壁细胞Figure3.10 Two conformation of circular double-stranded DNA.Figure3.11 Plasmid purification by the alkaline denaturation method.Alkaline denaturationFigure3.12 CsCl density gradient centrifugation.C

4、sCl density gradient centrifugationFigure3.13 Partial unwinding of the DNA double helix by EtBr intercalation between adjacent base parirs.EB如何与如何与DNA结合结合?Figure3.14Purification of plasmiod DNA by EtBr-CsCl density gradient centrifugation如何分离质粒如何分离质粒DNA？Figure3.15 Plasmid amplification Plasmid ampli

5、ficationInhibitor of protein synthesis: Chloramphenical, 12h, Figure3.17 Induction of a clts lysogen（溶原菌）（溶原菌） by transferring from 30 to 42Figure3.18 Achieving the right balance between culture age and inoculum size when preparing a sample of a non-lysogenic phage由于缺失修饰，大由于缺失修饰，大多数基因工程载体多数基因工程载体属于此类型。属于此类型。Figure3.19 Collection of phage particles by polyethylene（聚（聚乙烯）乙烯） glycol（乙二醇）（乙二醇）(PEG) precipitation Figure3.20 Purification of phage particles by CsCl density gradient centrifugation.