A MICROARRAY-BASED HIGH-THROUGHPUT GENOTYPING METHOD – THE TAGGED MICROARRAY MARKER (TAM) APPROACH

¹ Plant Research Unit, University of Dundee at SCRI, Invergowrie, DUNDEE DD2 5DA, United Kingdom

² Post-Genomics and Molecular Interactions Centre, School of Life Sciences, University of Dundee, DUNDEE DD1 5EH, UK

³ Scottish Crop Research Institute, Invergowrie, DUNDEE DD2 5DA, UK

⁴ Department of Crop Genetics, John Innes Centre, Colney Lane, NORWICH NR4 7UH, UK

⁵ Agrogene SA, 620 Rue Blaise Pascal ZI, 77550 Moissy Cramayel, FRANCE

A microarray-based method has been developed for scoring thousands of DNAs for a codominant molecular marker on a glass slide. The approach was developed to detect insertional polymorphism of transposons in pea and works well with indel polymorphisms in pea and single nucleotide polymorphism (SNP) markers in barley. 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. Flavell et al (2003) Nucleic Acids Res. 31: e115.