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Team:SDU-Denmark/Protocols/Electroporation
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(New page: =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...) |
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=Electroporation= | =Electroporation= | ||
Latest revision as of 10:06, 17 August 2009
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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.
- Thaw competent cells at room temperature and then keep on ice. 0.2cm cuvettes for electroporation are cooled on ice.
- 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.
- Place the cuvette in the electroporator and pulse once. Setup: Gene pulser: 25uF 2.5kV Pulse controller: 200ohm
- 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).
- Plate the transformed cells onto LA plates containing antibiotics (100 ug/ml ampicillin). Untransformed cells (negative control) should be plated with and without antibiotics.
