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Maintenance and Storage of DH5alpha E.coli cells
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Materials Required

 

  •  LB broth
  •  LB agar plates
  •  Micropipette and tips
  •  Bunsen burner
  •  Inoculation loop
  •  80% glycerol
  • -80°C freezer


Procedure


a)    To create a Glycerol Stock:



1.    Pick a single colony from a plate and grow an overnight in the LB broth.
2.    Add 0.5 ml of the overnight culture to 0.5 ml of 80% sterile glycerol in the microfuge tube.
3.    Vortex the mixture of overnight culture and glycerol slowly.
4.    Store the glycerol stock at -80 ºC.



b)    To pick the Culture from a Glycerol Stock:



1.    Take the microfuge tube from the -80 ºC freezer.
2.    Scrape off a portion of stored culture from the top of the frozen glycerol stock and streak it onto an LB agar plate.
3.    Invert the plates and incubate it for overnight incubation at 37ºC.
 

 

 

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