Analysis of two multiplexed short tandem repeat systems using capillary electrophoresis with multiwavelength fluorescence detection

Alice R. Isenberg, Ralph O. Allen, Kathleen M. Keys, Jill B. Smerick, Bruce Budowle, Bruce R. McCord

Research output: Contribution to journalArticle

26 Scopus citations

Abstract

A series of experiments was performed to analyze the utility of capillary electrophoresis (CE) with multiwavelength detection capabilities for multiplex typing of short tandem repeat loci. Characteristics of the sieving polymer, hydroxyethylcellulose, which affect resolution of single strand (ss) DNA fragments were examined. Additionally, the effects of denaturant in the polymer system, separation voltage, and analysis temperature were studied to ascertain their effects on DNA separations and capillary lifetime. The use of elevated run temperature (60°C) was found to improve sizing precision, to increase the lifetime of capillaries (100 runs or more per capillary), and to provide runtimes of under 20 min. Finally, 100 individual human DNA samples were typed successfully using CE. The average resolution obtained was 1.4 bases for a 200 base fragment with a standard deviation of sizing of 0.2 bases, allowing all alleles examined to be distinguished clearly.

Original languageEnglish
Pages (from-to)94-100
Number of pages7
JournalElectrophoresis
Volume19
Issue number1
DOIs
StatePublished - 1 Jan 1998

Keywords

  • Capillary electrophoresis
  • DNA
  • Fluorescence
  • Hydroxyethylcellulose
  • Polymerase chain reaction

Fingerprint Dive into the research topics of 'Analysis of two multiplexed short tandem repeat systems using capillary electrophoresis with multiwavelength fluorescence detection'. Together they form a unique fingerprint.

  • Cite this