Knight:Preparing stabs

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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&deg...)
Current revision (09:42, 2 July 2008) (view source)
 
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For long term storage of bacterial strains.  Useful for shipping strains.
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==Materials==
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*Stab agar
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**10 g nutrient broth
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**5 g NaCl
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**6 g agar
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**10 mg cysteine˙Cl
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**10 mg thymine
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**H<sub>2</sub>O to 1 L
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Autoclave to sterilize.
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==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
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#Seal tightly and store in the dark at room temperature (15-22&deg;C)
#Seal tightly and store in the dark at room temperature (15-22&deg;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
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[[Category:Protocol]] [[Category:Escherichia coli]]
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==References==
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<biblio>
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#ShortProtocols isbn=0-471-13781-2
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</biblio>

Current revision

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&dot;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. [ShortProtocols]
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