Knight:Preparing stabs

For long term storage of bacterial strains. Useful for shipping strains.

Materials

• Stab agar
  • 10 g nutrient broth
  • 5 g NaCl
  • 6 g agar
  • 10 mg cysteine˙Cl
  • 10 mg thymine
  • H₂O to 1 L

Autoclave to sterilize.

Procedure

1. Fill airtight, autoclavable vials with rubber or teflon caps 2/3 full of stab agar
2. Inoculate with a single colony repeatedly poking the inoculating loop into the agar
3. Incubate at 37°C for 8-12 hrs until cloudy tracks of bacteria become visible
4. Seal tightly and store in the dark at room temperature (15-22°C)
5. Revive by using inoculating loop to streak onto a new LB plate

References

1. ISBN:0-471-13781-2