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Position: Discussion forums > Bioscience biotechniques discussion forums > Molecular biology troubleshooting forum
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compatibility vector/competent cell
Posted by: frantzisca (IP Hidden, New member, 1)
Date: April 21, 2008 10:17AM
Hi,
could someone tell me something about possible incompatibility between some kind of vector and competent cells? I'm working with PBluescript KS+ (2,9 kb) containing a 6,5 kb DNA fragment, and I'm trying to ligate in it a 4,7 kb fragment. When I transform with OneShot TOP10 electrocomp E.Coli cells (following protocol conditions), the next day I find every time a patina over the entire plate, and any colonies, sometime 1-2, but negative. It is not contamination nor wrong Amp because I did the experiment by using the same stock of LB Amp plates but with different kind of cells (XL1Blue and TOP10 both chemically competents) and I never found this patina. I suspect some kind of incompatibility between the PBluescript (with a so large foreign DNA) and the OneShot TOP10 electrocomp E.Coli cells. I cannot find other explanation. Could you help me please? Frantzisca
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