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AlanFung:Protocol/Plasmid DNA amplification
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==Grow cells on LB-Plates== *Thaw transformation competent E.Coli cells on ice *Mix cells gently by lightly flicking the tube *Aliquot 50-100ul of cells into chilled 17x100mm polypropylene tubes *Unused cell may be frozen (For optimal recovery, refreeze cells in a dry ice/ ethanol bath prior to -70C storage *Add 2ul of diluted shRNA plasmid solution to 50ul cell suspension and gently swirl tube for a few second to mmix *Incubate on ice for 30mins *Place tubes in 42C water bath for 30 to 45 seconds without shaking *Place tube again on ice for 2 mins *Dilute transformation reactions to 1ml by addition of 900-950ul SOC medium *Shake the tube 200rpm for 60 minutes at 37C *Plate by spreading 5-200ul of cell transformation mixture on LB-Chloramphenicol plates containing appropriate antibiotic and incubate overnight at 37C until colonies appear
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