With the Promega pGEMT-easy cloning kit, I have trouble getting positive clones. I use electroporation instead of heat shock for transforming my JM109 cells. For this, I precipitate the ligation and resuspend it in water prior to electroporation. I wonder if this is the cause of the trouble. Has anyone ever tried electroporation with this kit? what are your experiences?
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perneseblue
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Unlimited ligation works!
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Posted 19 July 2010 - 06:16 PM
Are your JM109 cells electrocompetent cells? If so, I don't see why you should have any problems.
However the kit below comes with chemical competent cells.... which can't be used with an electroporation set up.
pGEMŪ-T and pGEMŪ-T
Easy Vector Systems
INSTRUCTIONS FOR USE OF PRODUCTS A1360, A1380,
A3600 AND A3610.
(I do hope I am looking at the right cloning kit)
However the kit below comes with chemical competent cells.... which can't be used with an electroporation set up.
pGEMŪ-T and pGEMŪ-T
Easy Vector Systems
INSTRUCTIONS FOR USE OF PRODUCTS A1360, A1380,
A3600 AND A3610.
(I do hope I am looking at the right cloning kit)
May your PCR products be long, your protocols short and your boss on holiday
