IGEM:IMPERIAL/2009/Transforming Competent Cells

Aims
To transform our electrocompetent E. coli with plasmids containing selected DNA/Parts/BioBricks.

Equipment

 * Desktop centrifuge
 * Electroporation machine
 * Electrocuvettes
 * Eppendorf tubes
 * P1000 and P200 Gilsons and tips

Reagents

 * XL-1 Blue competent cells (stored in the freezer)
 * LB
 * Plates with required antibiotic

Protocol

 * 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