Energy-transfer cassette labeling for capillary array electrophoresis short tandem repeat DNA fragment sizing

L. Berti, I. L. Medintz, J. Tom, R. A. Mathies

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Energy-transfer (ET) dye-labeled primers significantly improve fluorescent DNA detection because they permit excitation at a single common wavelength and they produce well separated and intense acceptor dye emission. Recently, a new ET cassette technology was developed [Berti, L. et al. (2001) Anal. Biochem. 292, 188-197] that can be used to label any PCR, sequencing, or other primer of interest. In this report we examine the utility of this ET cassette technology by labeling seven different short tandem repeat (STR) specific primers with each of the four ET cassettes and analyzing the PCR products generated on a MegaBACE-1000 capillary array electrophoresis system. More than 60 amplicons were generated and successfully analyzed with the ET cassette-labeled primers. Both forward and reverse primers were labeled for multiplex PCR amplification and analysis. Single base pair resolution was achieved with all four ET cassettes. This ET cassette-primer labeling procedure is ideally suited for creating four-color fluorescent ET primers for STR and other DNA assays where large numbers of different loci are analyzed including sequencing, genetic identification, gene mapping, loss of heterozygosity testing, and linkage analysis.

Original languageEnglish (US)
Pages (from-to)493-500
Number of pages8
JournalBioconjugate Chemistry
Volume12
Issue number4
DOIs
StatePublished - 2001

Fingerprint

Energy Transfer
Capillary Electrophoresis
Electrophoresis
Microsatellite Repeats
Energy transfer
Labeling
DNA
Coloring Agents
Dyes
Technology
Polymerase Chain Reaction
Loss of Heterozygosity
Chromosome Mapping
Multiplex Polymerase Chain Reaction
Base Pairing
Amplification
Labels
Assays
Color
Genes

ASJC Scopus subject areas

  • Chemistry(all)
  • Organic Chemistry
  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Energy-transfer cassette labeling for capillary array electrophoresis short tandem repeat DNA fragment sizing. / Berti, L.; Medintz, I. L.; Tom, J.; Mathies, R. A.

In: Bioconjugate Chemistry, Vol. 12, No. 4, 2001, p. 493-500.

Research output: Contribution to journalArticle

Berti, L. ; Medintz, I. L. ; Tom, J. ; Mathies, R. A. / Energy-transfer cassette labeling for capillary array electrophoresis short tandem repeat DNA fragment sizing. In: Bioconjugate Chemistry. 2001 ; Vol. 12, No. 4. pp. 493-500.
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