SREL DNA Lab
PFGE Methods

Five largemouth bass were collected by angling from L-Lake on the Savannah River Site in March 2000.  Blood was drawn from fish after they were measured, then the fish were released.  We tested storage buffers and methods according to the experimental design shown in the accompanying figure.

Dilute bass blood was embedded in agarose plugs to immobilize the DNA, then the red blood cells (now in plugs) were lysed to release the DNA.  These plugs were subsequently loaded into agarose gels and run in the pulsed field rig at low voltage for 16 hours.  Gels were stained with a dye that fluoresces under UV light and photographed using a digital imaging system.  Results were visually scored to estimate treatment effect.  Other variations of the technique were also tested and evaluated.

Once PFGE was satisfactorily refined, we tested it on eight other species of non-mammalian vertebrates (see Gallery).  We then collected 20 more bass in June 2000 from Langley Pond near Aiken, SC; their blood was analyzed for both DNA strand breakage and mercury content to see if there was some cause-effect correlation.