IGEM:IMPERIAL/2006/LabCalendar/2006-7-13

Electroporation

 * Raise voltage to about 2.2 to 2.3 V, the other knobs should already be set
 * Ensure the pulse controller is set at 200

Ensure:
 * No sample with high salt concentration, will cause to spark
 * Sample must be even across cuvette
 * Make sure the cuvette is dry!
 * You can use the cuvette several times (wash about 10 times with tap water and a few times with distilled water...place in 37C incubator to dry and set to autoclave for sterilisation)


 * Cool cuvettes (prechill) on ice before using
 * Always have a control to ensure that there is no noise when getting results


 * Competant bacteria are kept in -80C freezer down the corridor (bring your swipecard!)
 * Leave bacteria on ice
 * Make sure bacteria are thawed
 * Pipette all of the 50 uL aliquot of bacteria after mixing with a pipette, ensure that there are no bubbles in the cuvette!
 * If you get a spark, throw the curvette away and start again, clear the machine by pressing both buttons again until it beeps (releases charge)

After electroporation
 * Put 1 mL of LB (no antibiotic) pipette a few times and transfer to an eppendorf
 * Take to lab and spin for 10 seconds
 * Pippete off most of the liquid and resuspend
 * Transfer to agar plate and allow to grow