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

I. Kheterpal, J. R. Scherer, S. M. Clark, A. Radhakrishnan, J. Ju, C. L. Ginther, G. F. Sensabaugh, R. A. Mathies

Research output: Contribution to journalArticlepeer-review

107 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)1852-1859
Number of pages8
JournalELECTROPHORESIS
Volume17
Issue number12
DOIs
StatePublished - 1996

Keywords

  • Capillary electrophoresis
  • Confocal fluorescence detection
  • DNA sequencing
  • Energy transfer primers
  • Mitochondrial dna polymorphisms

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry

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