Knight:Preparing stabs: Difference between revisions
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(New page: #Fill airtight, autoclavable vials with rubber or teflon caps 2/3 full of stab agar #Inoculate with a single colony repeatedly poking the inoculating loop into the agar #Incubate at 37°...) |
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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 | |||
**H<sub>2</sub>O to 1 L | |||
Autoclave to sterilize. | |||
==Procedure== | |||
#Fill airtight, autoclavable vials with rubber or teflon caps 2/3 full of stab agar | #Fill airtight, autoclavable vials with rubber or teflon caps 2/3 full of stab agar | ||
#Inoculate with a single colony repeatedly poking the inoculating loop into the agar | #Inoculate with a single colony repeatedly poking the inoculating loop into the agar | ||
| Line 4: | Line 18: | ||
#Seal tightly and store in the dark at room temperature (15-22°C) | #Seal tightly and store in the dark at room temperature (15-22°C) | ||
#Revive by using inoculating loop to streak onto a new LB plate | #Revive by using inoculating loop to streak onto a new LB plate | ||
[[Category:Protocol]] [[Category:Escherichia coli]] | |||
==References== | |||
<biblio> | |||
#ShortProtocols isbn=0-471-13781-2 | |||
</biblio> | |||
Latest revision as of 06:42, 2 July 2008
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
- Fill airtight, autoclavable vials with rubber or teflon caps 2/3 full of stab agar
- Inoculate with a single colony repeatedly poking the inoculating loop into the agar
- Incubate at 37°C for 8-12 hrs until cloudy tracks of bacteria become visible
- Seal tightly and store in the dark at room temperature (15-22°C)
- Revive by using inoculating loop to streak onto a new LB plate
