hybing without shaking??

[Ferland Louis H.]

On Mon, 22 Apr 1996, Lely Chow wrote:

> Date: Mon, 22 Apr 1996 17:50:41 -0700
> From: Lely Chow <lelyc at u.washington.edu>
> To: methods at net.bio.net
> Subject: hybing without shaking??
> 
> 
> Hi,
> 
> I was doing an overnight hybe of some Northern and RNA dot blots in a
> shaking waterbath.  Something went wrong and the waterbath had stopped
> shaking when I came in the next morning.  Is shaking absolutely necessary
> for a good signal??  I had 4 blots in 50 mls of hybe solution in a small
> pyrex container.  There was just enough hybe to cover all of the blots and
> still allow for diffusion between blots.
> 
> Thanks in advance,
> 
> --Lely
> 
> 
> 
> 
> 
Lely, 

Expose your blots and you'll know!

In my experience, shaking does help getting a stronger and more uniform 
signal, but is not absolutely required. I know at least one lab where 
hybs are routinely done in a non-shaking waterbath, and they generate 
good results.


Dr. Louis H. Ferland
Centre de Recherche, Hotel-Dieu de Montreal
Dept de Nutrition, Universite de Montreal
Phone: (514) 843-2757     FAX: (514) 843-2719