Q: Southern Hyb times

Lee V. Madrid lmadrid at crcvms.unl.edu
Mon Oct 9 17:11:42 EST 1995

On Fri, 6 Oct 1995 scalzod at fred.fhcrc.org wrote:

> Hi,
>     I am wondering about the hybridization time for probing southern blots.  I 
> read times ranging from 6hrs to O/N.  What factors move the hyb time to 6 
> hours? Or less? I've heard rumour of rapid hybridization, but haven't been 
> able to confirm it.  
>     Our lab tends to load ~20ug genomic DNA and probe with ~3 million 
> cpm/ml hyb buffer and O/N hyb (mostly from tradition; it worked and it 
> continues to work).  We mostly use it to screen clones for integration 
> (whether a clone flpped or not).    
>      What's the shortest time you've probed, under what conditions and how did 
> it turn out?
>      thanks
We have had good experiences using Stratagenes (La Jolla, CA) "QuickHyb" 
Hybrdization buffer. According to the protocol one may hybridize for one 
hour or longer and it seems to work. Check it out.


Lee Madrid 
Pfizer Central Research
Molecular Biology
lmadrid at crcvms.unl.edu

"Just Clone It"


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