Capillary gel electrophoresis for rapid, high resolution DNA sequencing.
Capillary gel electrophoresis has been demonstrated for the separation and detection of DNA sequencing samples. Enzymatic dideoxy nucleotide chain termination was employed, using fluorescently tagged oligonucleotide primers and laser based on-column detection (limit of detection is 6,000 molecules per peak). Capillary gel separations were shown to be three times faster, with better resolution (2.4 x), and higher separation efficiency (5.4 x) than a conventional automated slab gel DNA sequencing instrument. Agreement of measured values for velocity, resolution and separation efficiency with theory, predicts further improvements will result from increased electric field strengths (higher voltages and shorter capillaries). Advantages of capillary gel electrophoresis for automatic DNA sequencing instruments and for genomic sequencing are discussed.
Swerdlow
H
H
Department of Human Genetics, Howard Hughes Medical Institute, University of Utah, Salt Lake City 84132.
Gesteland
R
R
D001483:Base Sequence; D004247:DNA / Q000235:genetics*; D004275:DNA, Ribosomal / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel / Q000295:instrumentation / Q000379:methods; D008969:Molecular Sequence Data; D012354:RNA, Transfer, His / Q000235:genetics; D012475:Salmonella / Q000235:genetics
D004275:DNA, Ribosomal; D012354:RNA, Transfer, His; D004247:DNA