UA Biophysics:Protocols:Stocks

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FIRST DAY

  1. Bacteria taken from either -80 tubes, or remaining stock and streaked onto antibiotic LB agar plates
  2. Grow plates overnight in hot room, collect early next day to get small colonies

SECOND DAY

  1. Select small single colony and add to 10ml LB (with antibiotics if resistance is in the strain) portioned into conical flask
  2. Place flask in hot room on shaker for overnight growth
  3. Collect flask early next day

THIRD DAY

  1. LB:Glycerol(50%) 3ml:2ml ratio
  2. Portion grown bacteria into eppendorfs (130ul each)
  3. Freeze bacteria in liquid nitrogen
  4. Store in -80C freezer

AMP AND KAN CONCENTRATION

1:1000 dilution of 100mg/ml amp 50mg/ml kan

Please, Do not hesitate to contact me if you have any question Juan Arias

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