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Unformatted text preview: -slowly mix cell suspension several times by pipetting and transfer into a 2 mm gap width cuvette -gently tap and/or flick cuvette several times until cell suspension settles at the bottom between the two metal plates -insert cuvette into electroporation apparatus and slide cuvette holder all the way back until cuvette is in contact with the electrodes -electroporation settings should be 2.5 kV, 25 uF, 200 -press and hold two red buttons simultaneously until tone sounds o time constant (displayed after electroporation) should be 4-5 msec -immediately resuspend cells in 1 ml room temperature LB and grow at 37C in rotator for 1-1.5 hours -transfer cells to a microcentrifuge tube, spin at 13,000rpm for 2 minutes and aspirate supernatant leaving the cell pellet in ~50 ul LB -resuspend cells in remaining 50 ul LB and plate on LB + antibiotic plate, incubate plates overnight at 37C o antibiotic should correspond to resistance conferred by plasmid...
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- Fall '10