Genotyping energy-transfer-cassette-labeled short-tandem-repeat amplicons with capillary array electrophoresis microchannel plates.
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.
Medintz
I L
IL
Department of Chemistry, University of California, Berkeley, CA 94720, USA.
Berti
L
L
Emrich
C A
CA
Tom
J
J
Scherer
J R
JR
Mathies
R A
RA
D002460:Cell Line; D019075:Electrophoresis, Capillary / Q000379:methods; D006801:Humans; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction; D013050:Spectrometry, Fluorescence