Sequencing gel haze near bottom

Ronald Meis camron at ubvms.cc.buffalo.edu
Sat Mar 26 16:11:00 EST 1994


I realize that this topic was discussed within the past year sometime
but do to recent moving, I didn't keep up to date, my apologies.

For some time I've been doing cycle sequencing with S35 internal
label with either the Circumvent and/or the fmol kit, and Long
Ranger gels. Reproduceably, I see a radioactive "haze" across all
lanes of any gel. This haze migrates slightly slower than the BB
dye at about 40 b. It is not a distinct band and covers the length
of upto 10 bases. Usually I can read the sequence from underneath
but not always. I've heard that this can happen with P32 internal
label but have never tried such.

Was there a consensus answer to the cause and/or cure of this on
the net? If I remeber correctly, it was proposed to be a thermal
breakdown product, but I didn't read if it was the label itself,
or a labeled compound in the reaction. How frequently is this
encountered? Again my apologies if this is a recent FAQ.

						Ronald Meis



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