McClean:Chloramphenicol media

Overview

Protocol for making chloramphenicol plates. This was adapted from Endy:Preparing_Antibiotic_Stocks. See also Chloramphenicol

CAUTION: Chloramphenicol is a suspected carcinogen.

Materials

1. LB1. See McClean:LB1
2. chloramphenicol (34mg/ml) stock solution in the -20C chest in 3007. See McClean:Chloramphenicol

Procedure

• Melt the LB1 in the microwave if it is solid. Start by microwaving it for 2 minutes, and then add 1 minutes intervals until it is liquid.
• Place the hot, liquid LB1 onto a shaker. This will let it cool without letting it solidify.
• When the LB1 is no longer painfully hot to the touch, add 500μL of the 34mg/mL chloramphenicol stock solution for each 100mL of LB1 for a working concentration of 170μg/mL. (NOTE- Addgene recommends a working concentration of 25 μg/mL of chloramphenicol. On 4/19/16 Cameron J. Stewart tested plates with 170μg/mL and 25μg/mL and found that only 170μg/mL was selective. This is probably because the chloramphenicol used is from 2006 and thus may be partially degraded.)
• Place the LB1+ chloramphenicol solution back on a shaker to for a little bit to ensure that it is well-mixed.
• Pour out the plates.

Notes

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Contact

*Cameron J. Stewart: April 19, 2016

or instead, discuss this protocol.