High-throughput SNP scoring with GAMMArrays: genomic analysis using multiplexed microsphere arrays

Green, Lance D. and Cai, Hong and Torney, David C. and Wood, Diane J. and Uribe-Romeo, Francisco J. and Kaderali, Lars and Nolan, John P. and White, P. Scott (2002) High-throughput SNP scoring with GAMMArrays: genomic analysis using multiplexed microsphere arrays.
Published In: Biomedical nanotechnology architectures and applications : 20 - 24 January 2002, San Jose, USA , SPIE. 4626 SPIE 2002, pp. 308-315.

Abstract

We have developed a SNP scoring platform, yielding high throughput, inexpensive assays. The basic platform uses fluorescently labeled DNA fragments bound to microspheres, which are analyzed using flow cytometry. SNP scoring is performed using minisequencing primers and fluorescently labeled dideoxynucleotides. Furthermore, multiplexed microspheres make it possible to score hundreds of SNPs simultaneously. Multiplexing, coupled with high throughput rates allow inexpensive scoring of several million SNPs/day. GAMMArrays use universal tags that consist of computer designed, unique DNA tails. These are incorporated into each primer, and the reverse-component is attached to a discrete population of microspheres in a multiplexed set. This enables simultaneous minisequencing of many SNPs in solution, followed by capture onto the appropriate microsphere for multiplexed analysis by flow cytometry. We present results from multiplexed SNP analyses of bacterial pathogens, and human mtDNA variation. Analytes are performed on PCR amplicons, each containing numerous SNPs scored simultaneously. In addition, these assays easily integrate into conventional liquid handling automation, and require no unique instrumentation for setup and analysis. Very high signal-to-noise ratios, ease of setup, flexibility in format and scale, and low cost make these assays extremely versatile and valuable tools for a wide variety of SNP scoring applications.


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Deposit Information:
ZAIK Number: zaik2004-471
Depositing User: Lars Kaderali
Date Deposited: 27 May 2004 00:00
Last Modified: 12 Jan 2012 11:38
URI: http://e-archive.informatik.uni-koeln.de/id/eprint/471