International Mammalian Genome Society

The 15th International Mouse Genome Conference (2001)


POSTER 158 - MICROARRAY-BASED SNP ANALYSIS OF MUTANT PROGENY DNA POOLS TO ESTABLISH CHROMOSOMAL LINKAGE

Mr Andrew Haynes
Medical Research Council
MRC UK Mouse Genome Centre & Mammalian Genetics Unit, Harwell
Oxford
OX11 0RD
UK

Co-Authors: Tymowska-Lalanne Z, Brown SDM, Denny P
Institutions: MRC UK Mouse Genome Centre and Mammalian Genetics Unit

The mapping of mutants arising from the ENU mutagenesis screen (Nolan, P.M., et al., Nat Genet. 2000 (4):440-3.) at Harwell is ongoing.  To date, 70 of the 170 confirmed inherited mutant phenotypes have either been assigned  to different locations in the mouse genome, or confirmed by non-complementation testing.  Currently the mapping analysis, to establish allele ratios, is conducted on pooled mutant progeny DNA, using an ABI 377 with a fluorescent microsatellite marker set.  Single nucleotide polymorphism (SNP) analysis may offer a more efficient and higher throughput means of mutant phenotype mapping.  We have investigated a microarray-based, SNP assay employing a single step, primer extension reaction incorporating fluorescent dNTPs (Pastinen, T., et al. Genome Res. 2000;10 1031-1042).  Using this method, allele specific oligos are printed on to isothiocyanate activated, silane coated slides, and SNPs (http://waldo.wi.mit.edu/SNP/mouse) tested for chromosomal linkage against pools of 30-40 N2 mutant affected progeny DNA.  Initial results indicate that it is possible to detect SNP linkage to a particular mutant phenotype, by quantification of fluorescence and comparison of allele ratios.


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