User:Vijay/Protocol/survival

Overview

Stationary phase survival test is used to analyse the survival ability of the pathogenic microbes after the stationary phase. This is done by incubating the culture for several weeks and plating at regular interval to determine the CFUs.

Materials

• Tryptic Soy Broth
• Tryptic Soy Agar

Procedure

1. Inoculate one single colony of the wild type and the mutant in 50ml of TSB (in a 500ml conical flask).
2. No antibiotics added in the medium
3. Incubate at 37C at 225 rpm for 2 weeks
4. At every 24hr interval,50ul of the culture was added with 450ul of TSB and used for serial dilution (till 10-8)
5. 50ul of each dilution is plated on TSA (containing antibiotic for mutant)
6. Spread the cultures and incubate at 37C
7. Count the colonies