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文献和实验for 20 minutes in 500 ml bottles in the RC5-B using the GS3 rotor. Resuspend the cell pellets in old media and transfer to two bottles, centrifuge as before, and decant the media. The cell pellets can be frozen at -70degC at this point. 3. Resuspend
1. Pick single colony and inoculate 250 ml of LB broth containing 100 m g/l ampicillin or appropriate antibiotic. Shake at 250 RPM overnight.2. Centrifuge cells in a Sovall GSA (250 ml)or SLA-3000 (500 ml) rotor at 5 k × g for 10 minutes
or appropriate antibiotic. Shake at 250 RPM overnight. 2. Centrifuge cells in a Sovall GSA (250 ml)or SLA-3000 (500 ml) rotor at 5 k × g for 10 minutes. 3. Resuspend cell pellet in 5 ml of GTE buffer (50 mM Glucose, 25 mM Tris-Cl, 10 mM EDTA, pH
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