RNase protections assays. Too much salt?

CAShug cashug at aol.com
Thu Mar 28 20:48:28 EST 1996


Hi,

Angela Hofstra
<Hofstra at aa.wl.com> wrote: 
Lately we've been having trouble with the last precipitation step in our 
RNase protection assays. We follow the procedure essentially as outlined 
in "Current Protocols in Molecular Biology".  After precipitating in 
ethanol we get lots of white precipitate, which I suspect is largely 
salt.  When we dissolve the pellet in loading buffer and load it on a 
vertical gel the sample rises.  

Has anyone encountered this before and what can we do about it?

As well I'd like to get people's opinions of the Maxiscript and HYBSpeed 
RPA kits from Ambion.

I can't help you with the salt problem but I can tell you that I've used
the Maxiscript and RPA II kits from Ambion--haven't yet used the HYBSpeed
kit. I've enjoyed great success so far--as long as I prepared the template
by PCR (I tacked on the T7 pol seq 5' to my antisense oligo) as
recommended by Ambion. If you want more info on the PCR template protocol,
email me. Good luck!
No affiliations with Ambion.



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