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Nucleic Acids Research, 2003, Vol. 31, No. 19 e115
© 2003 Oxford University Press

A microarray-based high throughput molecular marker genotyping method: the tagged microarray marker (TAM) approach

Andrew J. Flavell*, Viacheslav N. Bolshakov1, Allan Booth2, Runchun Jing, Joanne Russell2, T. H. Noel Ellis3 and Peter Isaac4

Plant Research Unit, University of Dundee at Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, UK, 1 Post-Genomics and Molecular Interactions Centre, School of Life Sciences, University of Dundee, Dundee DD1 5EH, UK, 2 Genome Dynamics Programme, Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, UK, 3 Department of Crop Genetics, John Innes Centre, Colney Lane, Norwich NR4 7UH, UK and 4 Agrogene SA, 620 Rue Blaise Pascal ZI, F-77550 Moissy Cramayel, France

*To whom correspondence should be addressed. Tel: +44 1382 562731; Fax: +44 1382 568587; Email: a.j.flavell{at}dundee.ac.uk

A microarray-based method has been developed for scoring thousands of DNAs for a co-dominant molecular marker on a glass slide. The approach was developed to detect insertional polymorphism of transposons and works well with single nucleotide polymorphism (SNP) markers. Biotin- terminated allele-specific PCR products are spotted unpurified onto streptavidin-coated glass slides and visualised by hybridisation of fluorescent detector oligonucleotides to tags attached to the allele- specific PCR primers. Two tagged primer oligonucleotides are used per locus and each tag is detected by hybridisation to a concatameric DNA probe labelled with multiple fluorochromes.


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