Screning E.coli transformants ...

Nick Theodorakis theodorn at gusun.georgetown.edu
Tue Feb 9 13:34:40 EST 1999


In article <bpmurray*STUFFER*-0802991245480001 at mac-daddy.ucsf.edu>,
  bpmurray*STUFFER*@socrates.ucsf.edu (Bernard P. Murray, PhD) wrote:

<snip>...
> this method to work?  I have a couple of really nasty
> ligations to perform at the moment and I am very tempted
> to try this as I know the desired clone is there somewhere
> (PCR) but many screeninge later I haven't found it
> (one more round and it will have to be colony hybridisation).
>      Thanks for any input,
>           Bernard
> --
> Bernard P. Murray, PhD
> Dept. Cell. Mol. Pharmacol., UCSF, San Francisco, USA
>

You have a higher tolerance for multiple minipreps than I have, Bernard.
Nowadays, if I can't find my clone in one pass, I go right to colony hybs.
Usually I find that something odd has happened, such as small inserts that
give blue colonies anyway.

I used to try a few rounds of screening with minis, then after I'd find the
clone with colony hybs, I'd think: that wasn't so hard, why didn't I do that
in the first place?

 _______________________________________________
| Nick Theodorakis                              |
| theodorn at gusun.georgetown.edu                 |
|_______________________________________________|

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