Integrated sample cleanup-capillary electrophoresis microchip for high-performance short tandem repeat genetic analysis.

Abstract

An integrated PCR sample cleanup and preconcentration process is developed for forensic short tandem repeat (STR) analysis using a streptavidin-modified photopolymerized capture gel injector for microchip capillary electrophoresis (microCE). PCR samples generated with one biotinylated primer and one fluorescent primer provide the input to the streptavidin-based affinity capture-microCE device. Monoplex PCR samples processed by the device exhibited approximately 10- to 50-fold increased fluorescence intensities, and DNA profiles generated using 9-plex STR samples displayed approximately 14- to 19-fold higher signal intensities compared to those analyzed using traditional cross injection. Complete STR profiles were obtained with as few as 25 copies of DNA template using the capture-microCE device. Four DNA samples with various degrees of degradation were also tested. Samples analyzed using the capture-microCE device resulted in a significant increase of successful allele detection. The ability of our capture-microCE device and method to remove contaminating ions, to concentrate the sample injection plug, and to eliminate electrokinetic injection bias provides a powerful approach for integrating sample cleanup with DNA separation.

DOI: 10.1021/ac8018685

Cite this paper

@article{Yeung2009IntegratedSC, title={Integrated sample cleanup-capillary electrophoresis microchip for high-performance short tandem repeat genetic analysis.}, author={Stephanie H I Yeung and Peng Liu and Nadia Del Bueno and Susan A. Greenspoon and Richard Mathies}, journal={Analytical chemistry}, year={2009}, volume={81 1}, pages={210-7} }