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=== Preparation of competent bacteria cells === | === Preparation of competent bacteria cells === | ||
− | + | #Grow a culture of bacteria in LB medium until it reaches OD600 = 0.5. | |
− | + | #Centrifuge cells for 10 minutes at 3500g at 4°C | |
− | + | #Resuspend the pellet slowly in 80mL of Tfb1 buffer. | |
− | + | #Centrifuge 5 min at 3500g at 4°C. | |
− | + | #Resuspend the pellet in 8 mL of Tbf2 buffer. | |
− | + | #Incubate 15 min in ice. | |
− | + | #Aliquot 200µL of cell suspension in sterile eppendorf tubes and conserve it at -80°C. |
Revision as of 13:12, 10 June 2016
Preparation of competent bacteria cells
- Grow a culture of bacteria in LB medium until it reaches OD600 = 0.5.
- Centrifuge cells for 10 minutes at 3500g at 4°C
- Resuspend the pellet slowly in 80mL of Tfb1 buffer.
- Centrifuge 5 min at 3500g at 4°C.
- Resuspend the pellet in 8 mL of Tbf2 buffer.
- Incubate 15 min in ice.
- Aliquot 200µL of cell suspension in sterile eppendorf tubes and conserve it at -80°C.