DNA ligation help
klevay at ukcc.uky.edu
Tue Jun 4 08:43:57 EST 1996
Here are my 2 cents:
Once I had very bad experience in cloning 12kb insert into Bluescript
with transformation into DH5a. Actually, all of purified plasmids
contained deletion of different size. Then I realised that the
products of leaking promotors? could be harmful for bacteria and lead
me into troubles. Interestingly, that DH5a doesn't have "lacIq" in its
genotype, i.e. overproduction of lac repressor protein, inhibiting
transcription from the lac promoter. Many other cells do have lacIq.
I transformed my ligation into JM101, got my construct from the first
hit and since then DH5a is on my black list (when I have tricky
U.of Kentucky, Lexington.
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