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transformation efficiency drops with ligation product


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#1 honey357

honey357

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Posted 24 April 2010 - 08:24 AM

i've been having a problem with my transformation efficiency dropping to nearly nothing when i transform with ligated product.

im using pRS426.. i've tested it by cutting it once with each restriction enzyme, religating vector and transforming which gave me normal results (lawn of colonies). its when i do a sequential digest of the vector and insert, followed by a ligation and transformation that i get 0 to 3 colonies.

i've checked the ligation reaction on a gel, and it is working fine.

any ideas?




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