Difference between revisions of "Team:Aix-Marseille/Experiments/Protocols"

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=== Preparation of competent bacteria cells ===
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= Protocols =
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== Protocol #1 : Preparation of competent bacteria cells ==
  
 
#Grow a culture of bacteria in LB medium until it reaches OD600 = 0.5.
 
#Grow a culture of bacteria in LB medium until it reaches OD600 = 0.5.

Revision as of 13:14, 10 June 2016

Protocols

Protocol #1 : Preparation of competent bacteria cells

  1. Grow a culture of bacteria in LB medium until it reaches OD600 = 0.5.
  2. Centrifuge cells for 10 minutes at 3500g at 4°C
  3. Resuspend the pellet slowly in 80mL of Tfb1 buffer.
  4. Centrifuge 5 min at 3500g at 4°C.
  5. Resuspend the pellet in 8 mL of Tbf2 buffer.
  6. Incubate 15 min in ice.
  7. Aliquot 200µL of cell suspension in sterile eppendorf tubes and conserve it at -80°C.