IGEM:IMPERIAL/2009/Transforming Competent Cells
To transform our electrocompetent E. coli with plasmids containing selected DNA/Parts/BioBricks.
- Desktop centrifuge
- Electroporation machine
- Eppendorf tubes
- P1000 and P200 Gilsons and tips
- XL-1 Blue competent cells (stored in the freezer)
- Plates with required antibiotic
- 1ul DNA into competent cell suspension. Pipette up and down.
- Take 50ul – put into electro-cuvette. Tap to ensure even distribution.
- Electroporate – make sure the machine is set to ‘bacteria’ on the mV setting. You should get a reading between 3.00 and 4.00 on the m/s read-out.
- Add 450ul LB to cuvette. Pipette up and down.
- Take 450ul and put into a new eppendorf tube. Put in the shaking incubator for 30mins.
- Centrifuge for 1minute.
- Discard supernatant.
- Plate out cells.
- Incubate overnight at 37°C