General Zamanian Lab recipes.

Ampicillin stock (100 mg/mL)

  1. Mix 2 g of ampicillin powder (Fisher AC611770250) into 20 mL of MilliQ H₂O.
  2. Vortex until the ampicillin is dissolved.
  3. Sterilize using a 50 mL syringe filter into a 50 mL conical tube.
  4. Aliquot 1 mL of the ampicillin solution into labeled 1.5 mL tubes and store at -20°C.

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Filtering and aliquoting should be performed using sterile technique in the hood.

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Kanamycin stock (50 mg/mL)

  1. Mix 500 mg of kanamycin powder (Fisher AC450810100) into 9 mL of MilliQ H₂O.
  2. Vortex until the kanamycin is dissolved.
  3. Aliquot 1 mL into labeled 1.5 mL tubes and store at -20°C.

LB Miller broth

  1. Mix the following:
Reagent Amount
Tryptone 10 g
Yeast extract 5 g
NaCl 10 g
dH₂O up to 1 L
  1. Autoclave on liquid cycle for 30 minutes.
  2. Add antibiotics after the media has cooled, if needed:
Antibiotic Stock Dilution Final Volume (per 500 mL media)
Ampicillin 100 mg/mL 1:1000 100 μg/mL 0.5 mL
Kanamycin 50 mg/mL 1:1000 50 μg/mL 0.5 mL
Chloramphenicol 50 mg/mL 1:3333 15 μg/mL 0.15 mL
  1. Store media with antibiotics at 4°C.

LB Miller agar

  1. Mix the following: