本科生《P》第五章目的基因制备与克隆策略概述.ppt

Chapter 6 估计基因组文库大小的经验（empirical formula）公式： N=ln(1-P)/ln(1-a/b) 选择哪种载体用于基因组文库构建？ Which vector is used to choose for construction of genetic library? 建立基因组文库的一般程序： 1．载体DNA片段的制备 载体DNA分离纯化 限制酶切 脱磷酸化反应  Homopolymer tailing: the enzyme terminal transferase is used to add homopolymers of dA, dT, dG or dC to a DNA molecule. Pst I vector CCCCCCCC-3’ 3’-CCCCCCCC Insert CTGCAGGGGGGGG-3’ G 3’-GGGGGGGGACGTC G Pst I site GGGGGGGGACGTC CTGCAGGGGGGGG GACGT TGCAG CCCCCCCC CCCCCCCC Pst I site Enzyme digestion Terminal transferase + dC Pst I digestion Terminal transferase + dG DNA ligase T