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
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