McClean: Carbenicillin Stock Solution
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.
- Carbenicllin (Novagent #69101)
- Sterile Mili-Q water
- Syringe (Becton-Dickinson #309653)
- 0.2 μm filters (Corning, Cat # 431229).
- Cryotubes (Nunc, Cat #375418)
- 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.)
|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|
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
- List troubleshooting tips here.
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Please sign your name to your note by adding '''*~~~~''': to the beginning of your tip.
- Megan N McClean 14:01, 20 July 2011 (EDT)
or instead, discuss this protocol.