AFLP Problem - Smearing
Elena Maria Dimuzio
emd15 at CORNELL.EDU
Mon Nov 8 10:17:28 EST 1999
I am experiencing problems with AFLPs. My reactions were working
beautifully last summer. I have just started doing them again (using
the same pre-amplified DNA) and I'm seeing the same basic patterns on
the gel in terms of major and minor bands, but the major bands are
smearing - instead of resolving into a sharp band, they leave a little
tail behind them in the gel. It looks like hair streaming back from
the band. The minor bands are also much less intense then they were
before. I know the template DNA I'm using is ok, and Life Tech sent me
a new kit which they've had no complaints about for the primers and PCR
buffer. I thought it was the gel, and have gotten new reagents for all
the gel components, but it doesn't seem to matter - I still have the
same problem. Has anyone encountered anything like this before?
Again, I have changed all the PAGE gel components with no success.
Another strange fact is that the AFLP ladder on the gels seems to run
just fine, a little bit blurry maybe but not big streaking like I see
in the sample lanes (even with the Life Tech pre-amplified DNA).
Thanks for your help.
emd15 at cornell.edu
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