Bands in all lanes [SOUTHERN]
Shin-ichiro HIRAGA
shiraga4 at bio.sci.osaka-u.ac.jp
Thu Nov 2 07:16:52 EST 1995
In article <DHE0L4.L3M at riker.neoucom.edu>, xzhang at riker.neoucom.edu
(Xiaolan Zhang) wrote:
>I am performing a genomic southern using 6 REs and DNA from 2 species. My
>probe is a 70bp fragment from the 5' end of my cDNA which I have labeled
>by random priming. The probe does not appear to contain repeat sequences.
>The DNA was digested well and transferred to nylon membranes.
>Hybridization at high stringency and exposure of X-ray film for 1 week was
>used. The blots are very clean.
>
>My results have been consistently puzzling. There are 8 bands ranging in
>size from 1 to 8 kb that appear in all lanes regardless of RE or specie.
>These bands vary in intensity but are about the same as the specific
>pattern over which they are superimposed.
>
>I think that if there was a contaminating DNA in the samples it would not
>yield the same bands after digestion with different REs. I tested my
>loading buffer for contamination by running just buffer and found none.
>There should not be so many introns covered by such a short probe. And
>they could not all happen to have the same RE sites.
>
>What could these bands be from and how can I solve this puzzle?
>
Hi,
Mnnnn, I cannot think of a good answer.
However, it seems that a there may be a problem in the electrophoresis step.
How about dye solution for electrophoresis? Isn't it contaminated?
Ah, this could not be any help. Sorry.
--
Shin-ichiro Hiraga
Graduate Student
Laboratory of Genetics
Faculty of Science
Osaka University
Toyonaka, Osaka 560
Japan
shiraga4 at bio.sci.osaka-u.ac.jp
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