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    • CommentAuthorkalayman
    • CommentTimeMar 30th 2010 edited

    This protocol explains how to: make LB plates for growing bacteria

    Written by: Jennifer Round

    5 g Tryptone
    2.5 g Yeast extract
    5 g NaCl

    Bring to 500 mls with Millipore water
    While mixing, prepare a 1-liter flask  with 7.5 g of  agar
    Adjust LB to pH 7.5 with dilute NaOH
    Pour liquid into flask with agar
    **ADD STIR BAR**
    Cover with aluminum foil and secure with autoclave tape
    Autoclave 30 min

    Allow to cool with slow mixing to about 50C (should be able to leave your hands on the glass without it hurting)
    Meanwhile, remove 1 sleeve of plates from the plastic and save the plastic
    Label the Plates according to which antibiotic you are adding (ampicillin is double black lines
    Add antibiotic and mix again for a few minutes (Ampicillin at 100 ug/ml, Kanamycin at 50 ug/ml, or Chloramphenicol at 25 ug/ml)
    Pour into agar plates, pouring just a little more than it takes to cover the bottom of the plate
    Leave at room temperature overnight to reduce condensation
    Store inverted in platic sleeves at 4C (refrigerator)