screening library

[Mayumi Yagi]

Hi Caroline,

300 bases should be long enough.  Have you tried random priming?  In my
experience, the probes come out much hotter, and I've used 200 bp PCR
products to screen lambda libraries successfully.  There are several kits
out (Boeringher-Mannheim and Amersham come to mind, but I'm sure there are
others as well).

Hope this helps.



On Tue, 4 Mar 1997, Caroline Szymeczek-Seay wrote:

> I'm having trouble screening a plasmid cDNA library.  My probe is only
> 300 bases long.  I'm trying to pull a longer clone out.  I'm afraid that
> my probe is not long enough, or not hot enough, to pull out a rare
> clone.  Has anyone used nick translation to label a probe that is only
> 300 bp long?  (PCR labelling doesn't give me high enough specific
> activity.)
> any thoughts on this?
> Caroline Seay
> 
> 

------------------------------------------------------------------------------
Mayumi Yagi, Ph.D.
Research (151)
Veterans Affairs Medical Center
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