Help! I made a mistake

David F. Spencer dspencer at
Fri Nov 7 11:53:50 EST 1997

In article <peng.36.190.34633C9E at>,
peng.36 at (p.J) wrote:

> Hi, neters:
>        I made a wrong SSC solution.  It sould be sodium chloride and
> citrate.  But I made it with sodium chloride and Tris base.  The worst thing 
> is that I used this SSC as a transfer buffer to transfer the DNA from 1% 
> agrose gel to membrane when I did Southern.  Now, I do not know what I should 
> do, trash the membrane or contine.  Any body have any suggestions?  Thanks a 
> lot.P. J

I wouldn't worry. I'm assuming you're using nylon rather than
nitrocellulose. What you've done is a quasi-alkaline transfer. I use the
standard alkaline transfer method which is with 0.4M NaOH alone, whereas
straight Tris (i.e. unneutralized) can't have a pH higher than about 11 (1M
Tris is about 11.2).  You may actually find that your transfer worked
better than usual!


David F. Spencer, PhD
Dept. Of Biochemistry
Dalhousie University
Halifax, Nova Scotia

dspencer at
dspencer at

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