Northern Rapid Transfer->References

Bradley Turner bsturner at MBCRR.HARVARD.EDU
Thu Apr 24 12:49:09 EST 1997


Dear Eric,

Yes it can be done, and it works very well for RNA.  I use it all the time
very successfully to transfer RNA to positively charged nylon
membranes (Hybond-N+, Amersham).  The main modification is to use either 50mM NaOH or 8mM NaOH/3M NaCl for
RNA instead of the 0.4M used for DNA

These blots also have the advantage of being able to be reprobed many
times.  I've reprobed up to at least five times

Downward northern alkaline blotting has been
described in the literature as follows:

REFERENCES:

One-hour downward capillary blotting of RNA at neutral pH
Analytical Biochemistry 221, 303-305, 1994
P Chomczynski, K Mackey

One-hour downward alkaline capillary transfer for blotting of DNA and RNA
Analytical Biochemistry 201, 134-139, 1992
P Chomczynski

Rapid downward alkaline blotting of RNA and DNA onto S&S NC and Nytran
membranes
Application Notes number 595, December 1992
Schleicher and Schuell , Inc.
Keene, NH 03431 USA
800-245-4024 phone
603-352-3810 phone
603-357-3627 fax
http://www.s-and-s.com

Alkaline Northern Blots: Transfer of RNA from Agarose gels to
Zeta-Probe (R) membrane in dilute NaOH
Molecular Biology Reports 1(3):1-4, November 1987 (BioRad Lab.
Pub.)
S Vrati, DA Mann, KC Reed
BioRad Laboratories
Hercules, CA 94547 USA
800-424-6723 phone 
510-741-1000 phone
800-879-2289 fax
http://www.bio-rad.com

Hybond-N+: Protocols for nucleic acid blotting and hybridization
Amersham Life Science
800-323-9750 phone
http://www.amersham.com/life/

I hope this helps,
Brad Turner


**************************************************************
		    Bradley Turner
	        Beth Israel Deaconess
                    Medical Center

Harvard Medical School		617-667-1215 phone
Division of Gastroenterology	617-667-2767 fax
Room Dana 536			bsturner at mbcrr.harvard.edu
330 Brookline Avenue		bturner at bidmc.harvard.edu
Boston, MA 02215, USA		turner at sprcore.bidmc.harvard.edu
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