UA Biophysics:Protocols:Chloramphenicol

Chloramphenicol

Chloramphenicol inhibits protein biosynthesis by targeting the peptidyl transferase center on the large ribosomal subunit. Bacteriostatic against gram-negative and gram-positive bacteria. Used in large-scale plasmid preparation of low to moderate copy number plasmids as well as in CAT assays.

STOCK SOLUTION

Chloramphenicol is commonly used in a range of 25 - 30 µg/mL, but can be used at an approximate range of 25 - 75 µg/mL dependent on the application. Stock solutions are typically at 34 mg/mL

PROTOCOL (STORAGE)

According to desired amount of antibiotics, calculate the amount in grams that has to be weighed.

For example, for 20mL of 25mg/mL of Chloramphenicol:

25mg/mL*20mL=500mg

1. Weigh the Chloranphenicol
2. Chloramphenicol has to be diluted in EtOH (molecular grade)
3. Divide in aliquots of 500μL
4. Store at -20 °C

Chloramphenicol is light sensitive. Keep tightly closed in light-resistant containers. Stock solutions should be stored at 2-8°C and are stable at 37°C for 5 days. Aqueous solutions are neutral and stable over a wide pH range, with 50% hydrolysis occurring after 290 days.

USAGE

For culture plates, allow media to cool to 55 °C before adding antibiotic to a final concentration of 35-50 μg/ml.

For Example, for 300 mL of media at a final concentration of 30µg/mL:

C_i V_i = C_f V_f

34mg/mL* V_i = 0,03mg/mL*300mL

V_i = 9mg/(34mg/mL)

V_i = 0,264 mL

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