best membrane for Northerns

cgriffin at wco.com cgriffin at wco.com
Thu Oct 23 01:41:03 EST 1997


Hello Alexander, 

There does seem to be a good deal concerning Hybond of late. Well as
someone who beta tested their "new formulation", I in fact had some
problems at first. BUT...easily addressed. I had to modify my protocol,
for Northerns, as follows. 

 Prior to Prehybridization I incubate the membrane in a salt solution
identical to that in the Pre and Hyb. solutions. Typically, I use 2XSSC in
Northern analysis, hence I incubate the membrane for say 15 min. at the
incubation temp. in 2XSSC, decant, and add Prehyb. soln., and that's it.
Mind you this is Hybond N...not Hybond-N+ with which I have no experience.


Also,  reduce your probe to say .5 to 1 x10e6 cpm per ml hybridization
buffer, if perchance you've been using higher concentrations. 

I have been told of late that the Magna membrane, available through Fisher
is pretty good, but I've no personal experience with it. Indeed the
suggestion was related to this whole issue of the often failing Hybond
complaints which are becoming fairly common of late. 

Lastly, my method works with the Hybond N of say 4-7 months ago, if they
have yet again reformulated...all bets are off. 

Hope it helps, 



In article <v01530501b0751dbb5a04@[129.127.90.149]>,
adobrovic at medicine.adelaide.edu.au (Alex Dobrovic) wrote:

> from recent discussions on the group it seems that hybond n+ is no longer
> giving good results, or is it just plain hybond? can anyone recommend
> anything that is working well at the moment? sensationally, would be even
> better
> 
> Alexander Dobrovic, Ph.D.
> Chief Scientist
> Department of Haematology-Oncology
> The Queen Elizabeth Hospital
> Woodville, SA 5011, Australia
> 
> Affiliate Senior Lecturer
> Department of Medicine
> University of Adelaide
> 
> Tel 61-8-8222 6884
> Fax 61-8-8222 6046



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