Help! Genomic probe gives smears!
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:
> 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.
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 Email:leach at mbcrr.harvard.edu
_|____ Dept. of Pharmacology Phone: (617) 638-5323
/ o / Boston Univ. School of Med. Fax: (617) 638-4329
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