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

(→‎Preparation of competent bacteria cells)
(→‎Preparation of competent bacteria cells)
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#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.
−
 
 
#Centrifuge cells for 10 minutes at 3500g at 4°C
 
#Centrifuge cells for 10 minutes at 3500g at 4°C
−
 
 
#Resuspend the pellet slowly in 80mL of Tfb1 buffer.
 
#Resuspend the pellet slowly in 80mL of Tfb1 buffer.
−
 
 
#Centrifuge 5 min at 3500g at 4°C.
 
#Centrifuge 5 min at 3500g at 4°C.
−
 
 
#Resuspend the pellet in 8 mL of Tbf2 buffer.
 
#Resuspend the pellet in 8 mL of Tbf2 buffer.
−
 
 
#Incubate 15 min in ice.
 
#Incubate 15 min in ice.
−
 
 
#Aliquot 200µL of cell suspension in sterile eppendorf tubes and conserve it at -80°C.
 
#Aliquot 200µL of cell suspension in sterile eppendorf tubes and conserve it at -80°C.

Revision as of 13:13, 10 June 2016

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.