Ultra-high throughout rotary capillary array electrophoresis scanner for fluorescent DNA sequencing and analysis

James R. Scherer, Indu Kheterpal, Arun Radhakrishnan, William W. Ja, Richard A. Mathies

Research output: Contribution to journalArticle

58 Scopus citations

Abstract

We have constructed a rotary confocal fluorescence scanner and capillary array electrophoresis system that is designed to analyze over 1000 DNA sequencing or fragment sizing separations in parallel. Capillaries are arranged around the surface of a cylinder and a rotating objective in the middle of the cylinder excites and collects fluorescence from labeled DNA fragments as they pass the capillary detection window. The capillaries are pressure-filled with a replaceable matrix and the samples are electrokinetically injected in parallel from a stainless steel microtiter plate at the cathode end. We demonstrate that the instrument is capable of producing four-color data from all capillaries at scan rate of 4 Hz (corresponding to a linear scan velocity of 121 cm/s). M13 sequencing data were obtained using a 128 capillary array mounted in half of the first quadrant of the scanner. In this initial run, read lengths greater than 500 bases were obtained in over 60% of the capillaries.

Original languageEnglish (US)
Pages (from-to)1508-1517
Number of pages10
JournalElectrophoresis
Volume20
Issue number7
DOIs
Publication statusPublished - 1999

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Keywords

  • Capillary electrophoresis
  • Confocal scanning
  • DNA sequencing
  • Fluorescence detection

ASJC Scopus subject areas

  • Clinical Biochemistry

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