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