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
- Carbenicllin (Novagent #69101)
- Sterile Mili-Q water
- Syringe (Becton-Dickinson #309653)
- 0.2 μm filters (Corning, Cat # 431229).
- 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
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
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References
Contact
- Megan N McClean 14:01, 20 July 2011 (EDT)
or instead, discuss this protocol.