UA Biophysics:Protocols:Chloramphenicol

Chloramphenicol 25 mg/ml

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.

Materials

• Chloramphenicol
• Ethanol
• Syringe filter
• Syringe 50 ml

Protocol

1. According to desired amount of antibiotics, calculate the amount in grams that has to be weighed. For example, for 10mL of 25mg/mL of Ampicillin: 25mg/mL*10mL=250mg
2. Weigh the Chloranphenicol
3. Chloramphenicol has to be diluted in EtOH (molecular grade)
4. Filter the solution using a 0.2 µm syringe filter.
5. Divide in aliquots of 500μL
6. 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.

Return to Protocols