McClean: Carbenicillin Stock Solution

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Contents

Overview

Carbenicillin stock solution for making LB + Carb media for growing e. coli with AMP-resistant plasmids. Stocks are stored in -80°C freezer for long-term storage and moved to the 4°C (pink box) for working stocks.

Materials

  1. Carbenicllin (Novagent #69101)
  2. Sterile Mili-Q water
  3. Syringe (Becton-Dickinson #309653)
  4. 0.2 μm filters (Corning, Cat # 431229).
  5. Cryotubes (Nunc, Cat #375418)

Procedure

  • Dissolve 5g of Carbenicillin in 50ml of sterile water.
  • Syringe filter using 0.2 um filters
  • Divide into aliquots in cryotubes
  • Store at -80°C until first use. Then please take one vial out quickly at a time, label it with (1) 100 mg/ml Carb, (2) the date of the day that you took it out, and (3) your initials. Then keep the labeled tube in the common antibiotic stock in the 4°C fridge.
  • DO NOT put your used tubes back into the -80°C stock.
  • For selective media:
    • Add 0.5ml of the stock for 1 liter of LB+Carb broth (50 μg/ml final conc.)
    • Add 1ml of the stock for 1 liter of LB+Carb plates (100 μg/ml final conc.)

Handy Table

100mg/ml Carb LB+CARB BROTH 100 mg/ml Carb LB+CARB Plate
500 µl1,000 ml1000 µl1,000 ml
250 µl500 ml500 µl500 ml
125 µl250 ml250 µl250 ml
100 µl200 ml200 µl200 ml
63 µl125 ml125 µl125 ml
50 µl100 ml100 µl100 ml
25 µl50 ml50 µl50 ml
13 µl25 ml25 µl25 ml
5 µl10 ml10 µl10 ml

Notes

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References

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