Team:SDU-Denmark/Protocols/Electroporation

From 2009.igem.org

Home | Background | Project | Parts | Team

Diary | Protocols | Downloads | Brainstorm

Electroporation

This protocol is very useful compared to standard-transformation, because it uses the power of ZAP! Requires less raw material than a normal transformation and is faster.

  1. Thaw competent cells at room temperature and then keep on ice. 0.2cm cuvettes for electroporation are cooled on ice.
  2. For each transformation, transfer 40ul cells to a 1.5ml Eppendorf tube and add 1.5ul plasmid from distribution plates. Mix well and transfer to the bottom of a cuvette without making air bubbles. Keep on ice for 5min.
  3. Place the cuvette in the electroporator and pulse once. Setup: Gene pulser: 25uF 2.5kV Pulse controller: 200ohm
  4. Immediately add 1ml SOC medium to the cuvette, mix well by pipetting, and transfer to a 1.5ml Eppendorf tube and incubate immediately at 37˚C for 1h on shaker (500 rpm).
  5. Plate the transformed cells onto LA plates containing antibiotics (100 ug/ml ampicillin). Untransformed cells (negative control) should be plated with and without antibiotics.