Quick Change

[Charan]

Hi,
I am having issues with Quick change mutagenesis.
I am trying to intrduce a single base change in a gene cloned into a plasmid (5Kb). My oligos are 37 bp long with 18 bp homology on either side.
I am following the protocol as per the manual but only variation is that
a) My oligos are not PAGE purified
b) I donot use the super competent cells for transformation. I make my own(chemically competent).
At the end of it I get NO colonies.
Is my reaction not working or my transformation?????????/
In either case what would you recomend???????/
PL. chip in with idea(s). Any suggession(s) wil be appreciated.

[canalon]

At least you should have a transformation control, is it working? If not The problem is coming from your cells, if it does, from your mutagenesis, but I am not familiar with thi sprotocol, so I can't help...