McClean: Carbenicillin Stock Solution

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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Contact

• Megan N McClean 14:01, 20 July 2011 (EDT)

or instead, discuss this protocol.