Plant & Animal Genomes XI Conference

January 11-15, 2003
Town & Country Convention Center
San Diego, CA

Poster: High-throughput Methods
            

ULTRA-HIGH THROUGHPUT MULTIPLEX SNP GENOTYPING USING SNPWAVE

Keygene N.V., Agro Business Park 90, P.O. Box 216, 6700 AE Wageningen, The Netherlands

Large-scale genotyping of single nucleotide polymorphisms (SNP) is important for pharmaco-genomics applications and for marker-assisted selection in plant and animal breeding programs. Since a large diversity of economical platforms for SNP detection exist, the main cost component is preparation of the detection mixture. Thus, in order to reduce the cost per datapoint, efficient multiplex amplification and/or miniaturization of reaction volumes are needed. We present SNPWave^TM as an ultra-high throughput technology for multiplex amplification and detection of SNP in the genome. SNPWave^TM reduces the cost per assay dramatically due to the combination of multiplex allele-discrimination for 10 SNP loci in a single reaction. This is followed by multiplex detection using a (capillary) sequencer, such as the MegaBACE, using multiple fluorescent labels and multiple injections per run. The latter is possible because all amplification products fit within a narrow size window. Over 23,000 co-dominantly scored SNP can so be detected in a single run with 96 capillaries, 4 labels, 10 SNP loci per reaction and 6 repeated injections. SNPWave^TM can also be used to detect interesting target sequences such as introgression segments or transgenes, or a combination thereof with SNP. The SNPWave^TM technology will be presented using tomato as a model system. Application for trademark registration for SNPWave^TM has been filed by Keygene N.V.. MegaBACE is a trademark of Amersham BioSciences.