DNA sequencing using a four-color confocal fluorescence capillary array scanner

Electrophoresis. 1996 Dec;17(12):1852-9. doi: 10.1002/elps.1150171209.

Abstract

The design, construction and operation of a four-color capillary array electrophoresis scanner are presented. The use of sensitive energy transfer primers facilitates four-color detection of the DNA sequencing fragments following excitation at a single laser wavelength (488 nm). This scanner collects fluorescence data from up to 25 capillaries in parallel. The resulting four-color image files are automatically reduced to four-color line plots, and a base-calling program (Sax) is used to call the sequence. The performance of this system for DNA sequencing is demonstrated by examining twelve different motifs of the hypervariable region I of human mitochondrial (mt) DNA obtained from a Sierra Leone population.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Base Sequence
  • DNA, Mitochondrial / chemistry*
  • Electrophoresis, Capillary / instrumentation*
  • Electrophoresis, Capillary / methods
  • Humans
  • Molecular Sequence Data
  • Sequence Analysis, DNA / instrumentation*
  • Sequence Analysis, DNA / methods
  • Sierra Leone

Substances

  • DNA, Mitochondrial