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 e xpanding needs of studies of short tandem repeats (STRs), we demonstrated h igh-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 electrophoresi s (CAE) microchannel plate system. Four-color detection was accomplished wi th a laser-excited confocal fluorescence rotary scanner. Results: Multiplex STR analysis with single base-pair resolution was demons trated on denaturing polyacrylamide gel media. The high-throughput multiple x capabilities of this genetic analysis platform were demonstrated by the s imultaneous separation of STR amplicons representing 122 samples in ninety- six 5.5-cm-long channels in < 8 min. Sizing values obtained for these ampli cons on the CAE microchannel plate were comparable to those measured on a c onventional commercial CAE instrument and exhibit < 1% sizing variance. Conclusions: Energy-transfer-cassette labeling and microfabricated CAE micr ochannel plates allow high-performance multiplex STR analyses. (C) 2001 Ame rican Association for Clinical Chemistry.