Identification of DNA molecules by pre-column hybridization using capillary electrophoresis

Abstract In this paper, we demonstrate the feasibility of using gel-filled capillary electrophoresis to perform Southern blotting with on-line detection and to study parameters that affect the hybridization of DNA molecules in solution. A fluorescence-tagged oligonucleotide was used as the probe for hybridization in solution with complementary DNA molecules prior to electrophoresis. The reaction mixture was subject to capillary electrophoresis in a polyacrylamide gel-filled capillary, and the hybridized species was identified on-line by UV absorption or laser-induced fluorescence detection. The effects of probe concentration and annealing temperature were studied.