Help! Genomic probe gives smears!

Martin Leach leach at mbcrr.harvard.edu
Mon Feb 14 19:17:47 EST 1994


In article <1994Feb9.215355.20979 at kakwa.ucs.ualberta.ca>,
eatkinso at gpu.srv.ualberta.ca (Eric Atkinson) wrote:

 It's been 
> suggested that the most probable cause for this genomic probe not working is
> that it contains repetetive elements.   Does 
> anybody out there have any suggestions of how I might be able to use my originalprobe by somehow blocking the repetetive elements of the filter DNA?  Would using sonicated mouse DNA as a blocking agent in my prehyb. work?  I've never
> heard of this, but somebody else has suggested it.

Dear Eric,

I was looking at RFLP's with a human genomic probe on human genomic blots.
The probe had repetitive elements and i assume could have produced a smear
such that you have seen.  We used to add sonicated human DNA to the prehyb
and hyb to block out the smear and it worked.

The amount of DNA to use depends on how repetitive the element is.

If you need to know what we used in the human blots drop me a line and i
will look it up.

Martin Leach

-- 

.....          Martin Leach                Email:leach at mbcrr.harvard.edu 
   _|____      Dept. of Pharmacology       Phone: (617) 638-5323        
   / o  /      Boston Univ. School of Med. Fax:   (617) 638-4329         
 _/  |-/__==/  80 E. Concord St. (L603)
(BULLDOZER) \_ Boston MA 02118            "Not the old underpants on your
               USA                           head.....WIBBLE" -BLACKADDER  



More information about the Methods mailing list