alkali vs non-alkali transfers

Eric Lader lader at ambion.com
Mon Jul 28 20:05:04 EST 1997


On 18 Jul 1997 13:10:20 GMT, Ross S Feldberg
<rfeldber at emerald.tufts.edu> wrote:

>Hi;
>
> I am setting up Northern blots and was wondering about optimal transfer
>conditions. We are using downward blotting onto charged nylon filters with
>an alkaline transfer solution. The protocol we are following calls for
>only a 60 min transfer in order to minimize hydrolysis of the RNA.
>However, I am not sure if we are getting good transfer in this time frame. 
>
see if you can uv shadow the RNA on the membrane to check transfer.

Ambion uses an alkali blotting onto charged nylon. Check out our
upside down transfer (speeds the transfer time, no gel compression).
Critical to pour thin gels and limit transfer to two hours or less. 

Gives several fold higher signal. Keep at it. Also think about RNA
probes.



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