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Electroporation not working, need help

Electroporation not working, need help - Molecular Cloning Forum

Electroporation not working, need help - DNA cloning forum. Discuss the cloning of plasmids, vectors, DNA, RNA, cDNA, proteins, and other molecules for expression, sub-cloning, and other methods. Also discuss digestion, ligation, transformation, plasmid preparation, cDNA library screening, and competent cells here.


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Old 12-22-2008, 10:44 PM
Pipette Filler
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Default Electroporation not working, need help



I am trying to insert the plasmid using electroporation in gram negetive bacteria ( many of my lab mates have done in past).. Its only me. My electro-competent cells are fine as when I spead them on regular LB plate (no antibiotic), they are growing..

Now, I use single pulse, Use SOC as nutrient medium after giving pulse for recovery. My recovery time is 1hr. I use 0.1cm gap cuvette.

what steps can be done differently to trouble shoot? should i give longer pulse/ double pulse....
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