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文献和实验细胞裂解液(Cell Lysis Solution)的配置方法
1.1M Tris 溶液的配置 取Tris242.2g,加ddH2O至1600ml,加热溶解。 2.1M Tris-HCL Ph=8.0溶液的配置 取 1M Tris 溶液160ml用分析纯盐酸调至Ph=8.0(需浓盐酸约8.5ml),加ddH2O定容至200ml,高压灭菌备用。 3. 0.5M EDTA溶液的配置 称EDTA-Na2 37.2g 先用140ml ddH2O溶解,加入14ml NaOH(10M)使EDTA-Na2溶解,再用NaOH(10M)溶液调至Ph
9M Urea,2.5mM EDTA,2.5mM EGTA,1% DTE,4% CHAPS make 10ml and aliquot 10x1ml,freeze at -70℃ Lysate preparation wash the cells 2x with PBS wash the cells 1x with 10mM Tris,250mM Sucrose lyse the cells with 100 – 350μl of urea lysis buffer
Miniprep of Plasmid DNA (Alkaline Lysis Method)
of culture into an Eppendorf tube. Spin for 30 seconds to pellet cells. 3. Remove the medium by pipet or aspiration, leaving the pellet as dry as possible. 4. Resuspend pellet in 100 ul of ice-cold GTE solution by gently pipetting up and down. Let
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