Genotyping energy-transfer-cassette-labeled short-tandem-repeat amplicons with capillary array electrophoresis microchannel plates

I. L. Medintz, L. Berti, C. A. Emrich, J. Tom, J. R. Scherer, R. A. Mathies

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Background: Genetic analysis of microsatellite DNA is a powerful tool used in linkage analysis, gene mapping, and clinical diagnosis. To address the expanding needs of studies of short tandem repeats (STRs), we demonstrated high-performance STR analysis on a high-throughput microchannel plate-based platform. Methods: Energy-transfer-cassette-labeled STR amplicons were separated and typed on a microfabricated 96-channel radial capillary array electrophoresis (CAE) microchannel plate system. Four-color detection was accomplished with a laser-excited confocal fluorescence rotary scanner. Results: Multiplex STR analysis with single base-pair resolution was demonstrated on denaturing polyacrylamide gel media. The high-throughput multiplex capabilities of this genetic analysis platform were demonstrated by the simultaneous separation of STR amplicons representing 122 samples in ninety-six 5.5-cm-long channels in <8 min. Sizing values obtained for these amplicons on the CAE microchannel plate were comparable to those measured on a conventional commercial CAE instrument and exhibit <1% sizing variance. Conclusions: Energy-transfer-cassette labeling and microfabricated CAE microchannel plates allow high-performance multiplex STR analyses.

Original languageEnglish (US)
Pages (from-to)1614-1621
Number of pages8
JournalClinical Chemistry
Volume47
Issue number9
StatePublished - 2001

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Energy Transfer
Capillary Electrophoresis
Microchannels
Electrophoresis
Microsatellite Repeats
Energy transfer
Throughput
Chromosome Mapping
Base Pairing
Labeling
Lasers
Color
Genes
Fluorescence
DNA

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Medintz, I. L., Berti, L., Emrich, C. A., Tom, J., Scherer, J. R., & Mathies, R. A. (2001). Genotyping energy-transfer-cassette-labeled short-tandem-repeat amplicons with capillary array electrophoresis microchannel plates. Clinical Chemistry, 47(9), 1614-1621.

Genotyping energy-transfer-cassette-labeled short-tandem-repeat amplicons with capillary array electrophoresis microchannel plates. / Medintz, I. L.; Berti, L.; Emrich, C. A.; Tom, J.; Scherer, J. R.; Mathies, R. A.

In: Clinical Chemistry, Vol. 47, No. 9, 2001, p. 1614-1621.

Research output: Contribution to journalArticle

Medintz, IL, Berti, L, Emrich, CA, Tom, J, Scherer, JR & Mathies, RA 2001, 'Genotyping energy-transfer-cassette-labeled short-tandem-repeat amplicons with capillary array electrophoresis microchannel plates', Clinical Chemistry, vol. 47, no. 9, pp. 1614-1621.
Medintz, I. L. ; Berti, L. ; Emrich, C. A. ; Tom, J. ; Scherer, J. R. ; Mathies, R. A. / Genotyping energy-transfer-cassette-labeled short-tandem-repeat amplicons with capillary array electrophoresis microchannel plates. In: Clinical Chemistry. 2001 ; Vol. 47, No. 9. pp. 1614-1621.
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AU - Medintz, I. L.

AU - Berti, L.

AU - Emrich, C. A.

AU - Tom, J.

AU - Scherer, J. R.

AU - Mathies, R. A.

PY - 2001

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N2 - Background: Genetic analysis of microsatellite DNA is a powerful tool used in linkage analysis, gene mapping, and clinical diagnosis. To address the expanding needs of studies of short tandem repeats (STRs), we demonstrated high-performance STR analysis on a high-throughput microchannel plate-based platform. Methods: Energy-transfer-cassette-labeled STR amplicons were separated and typed on a microfabricated 96-channel radial capillary array electrophoresis (CAE) microchannel plate system. Four-color detection was accomplished with a laser-excited confocal fluorescence rotary scanner. Results: Multiplex STR analysis with single base-pair resolution was demonstrated on denaturing polyacrylamide gel media. The high-throughput multiplex capabilities of this genetic analysis platform were demonstrated by the simultaneous separation of STR amplicons representing 122 samples in ninety-six 5.5-cm-long channels in <8 min. Sizing values obtained for these amplicons on the CAE microchannel plate were comparable to those measured on a conventional commercial CAE instrument and exhibit <1% sizing variance. Conclusions: Energy-transfer-cassette labeling and microfabricated CAE microchannel plates allow high-performance multiplex STR analyses.

AB - Background: Genetic analysis of microsatellite DNA is a powerful tool used in linkage analysis, gene mapping, and clinical diagnosis. To address the expanding needs of studies of short tandem repeats (STRs), we demonstrated high-performance STR analysis on a high-throughput microchannel plate-based platform. Methods: Energy-transfer-cassette-labeled STR amplicons were separated and typed on a microfabricated 96-channel radial capillary array electrophoresis (CAE) microchannel plate system. Four-color detection was accomplished with a laser-excited confocal fluorescence rotary scanner. Results: Multiplex STR analysis with single base-pair resolution was demonstrated on denaturing polyacrylamide gel media. The high-throughput multiplex capabilities of this genetic analysis platform were demonstrated by the simultaneous separation of STR amplicons representing 122 samples in ninety-six 5.5-cm-long channels in <8 min. Sizing values obtained for these amplicons on the CAE microchannel plate were comparable to those measured on a conventional commercial CAE instrument and exhibit <1% sizing variance. Conclusions: Energy-transfer-cassette labeling and microfabricated CAE microchannel plates allow high-performance multiplex STR analyses.

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