Low Fluorescence DNA Gels

Eugenie L. Harris "jean.harris" at stonebow.otago.ac.nz
Sat Aug 16 02:18:37 EST 1997


Hi Bryan and Frank,

I concur with Frank that most people use far too much EtBr.  But I'm
going to go one better and say all this staining and destraining
business is a waste of time.  We use Metaphor agarose for microsatellite
typing and save a lot of money by reusing the gels up to 10 X in a
simple load and run, load and run strategy.  When we run a gel for the
first time we usually put EtBr in the gel before pouring it.  After that
we just put a mimimal amount of EtBr in the bottom tank.  The DNA
migrates to the anode.  The ethidium migrates to the cathode.  They
meet, the bands are stained, the photo is taken, the buffer is changed
and the gel is loaded and run again.  Since we use minimal EtBr,
background is very low.  The EtBr concentration in the bottom tank is
roughly 0.03 ng/ml.  (I'm not quite sure on that amount since I can't
remember the volume in the lower tank and am guessing it.)

Regards,
Jean



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