Is a 15 na oligo ok for colony hybridization?

Tom Chappell t.chappell at ucl.ac.uk
Mon Jan 26 11:52:15 EST 1998


In article <6aav58$svk$1 at nntp.ucs.ubc.ca>, rewhlab at cbdn.ca (REWHLAB) wrote:

>Hi,
>I am trying to screen for the insertion of 15 bases into a gene that is cloned 
>in a pUC based vector.  I do not have experience using oligos as probes for 
>colony blots; does anyone know if a 32P-labelled 15 base oligo will work for 
>screening colony blots?  Or is this oligo too small?
>
>Thanks,
>REW
>rewhlab at cbdn.ca

I've cloned a gene from a yeast genomic cosmid library with a 17mer,
basically you need to know the Tm. You'll be doing low temp hybridizations
(< RT) and you should go with TMAC washes.  Personally, I'd come up with a
PCR screen--unless there is something weird about what you are trying to
do, your worst case scenerio is screening a few hundred colonies by PCR.
This will be easier than getting the colony blots to work.

-- 
Tom Chappell
MRC Laboratory for Molecular Cell Biology
University College London

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