McClean: Carbenicillin Stock Solution

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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 µl 1,000 ml 1000 µl 1,000 ml
250 µl 500 ml 500 µl 500 ml
125 µl 250 ml 250 µl 250 ml
100 µl 200 ml 200 µl 200 ml
63 µl 125 ml 125 µl 125 ml
50 µl 100 ml 100 µl 100 ml
25 µl 50 ml 50 µl 50 ml
13 µl 25 ml 25 µl 25 ml
5 µl 10 ml 10 µl 10 ml

Notes

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References

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