Difference between revisions of "Knight:Preparing stabs"

From OpenWetWare
Jump to: navigation, search
(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...)
 
 
Line 1: Line 1:
 +
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&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
 +
 +
[[Category:Protocol]] [[Category:Escherichia coli]]
 +
 +
==References==
 +
<biblio>
 +
#ShortProtocols isbn=0-471-13781-2
 +
</biblio>

Latest revision as of 05:42, 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&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]