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文献和实验Transformation of E. coli by Electroporation
ml of SOB (without magnesium) in a 1 liter flask. Grow for 2 to 3 hours with vigorous aeration at 37EC until the cells reach an OD550 = 0.8. 3. Harvest cells by centrifugation at 5000 RPM (2,600 g) in a GSA rotor for 10 min. (Make sure you use
培养基中,37℃振摇过夜。 (2)接种25ml过夜培养物于500ml LB培养基,37℃振摇,至OD600 达到0.4. (3)迅速将培养物置于冰浴中30min,至充分冷却。 (4)将菌液转移至预冷的离心管中,4℃下以2500r/min离心15 min,弃培养液,回收细胞。 (5)以10ml预冷的10%甘油洗涤沉淀,低温离心,共两次。 (6)加约1ml(适量)预冷的10%甘油重悬沉淀,稀释悬液100倍,测量OD600,至稀释浓度为2-3×1010个细胞/ ml (1.0 OD600约
of cells that receives no plasmid DNA at all. Store the tubes on ice for 30 minutes. Transfer the tubes to a rack placed in a preheated 42ºC circulating water bath. Store the tubes in the rack for exactly 90 seconds. Do not shake
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