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Simultaneous genetic typing from multiple short tandem repeat loci using a 96‐capillary array electrophoresis system
Author(s) -
Gao Qiufeng,
Pang Homing,
Yeung Edward S.
Publication year - 1999
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(19990601)20:7<1518::aid-elps1518>3.0.co;2-5
Subject(s) - capillary electrophoresis , genotyping , typing , polymerase chain reaction , microsatellite , chromatography , tandem , genomic dna , locus (genetics) , biology , microbiology and biotechnology , analytical chemistry (journal) , allele , genetics , materials science , genotype , dna , chemistry , gene , composite material
Short tandem repeat (STR) markers are highly polymorphic and widely used in human identification and genetic mapping. We demonstrate fast and reliable genotyping based on the four STR loci vWF, THO1, TPOX, CSF1PO by multiple‐capillary array electrophoresis. Extracted human genomic DNA was amplified by polymerase chain reaction (PCR). The PCR products were mixed with pooled allelic ladder as an absolute standard and coinjected from a 96‐vial tray. Separations were performed in polyvinylpyrrolidone (PVP) sieving matrix with a one‐hour turnaround time, with no degradation over 27 runs. Simultaneous one‐color laser‐induced fluorescence detection was achieved by using a charge‐coupled device (CCD) camera. The allele peaks for the unknown sample were identified by comparing the normalized peak intensities of the mixtures to those of the pooled ladder by using a straightforward algorithm. An extremely high level of confidence in matching the bands was indicated with negligible crosstalk (< 0.89%) between adjacent capillaries. This scheme is applicable for STR genotyping with high resolution, high speed and high throughput.