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Microsatellite genotyping by primer extension and MALDI-ToF mass spectrometry

Paris, M. and Jones, M.G.K.ORCID: 0000-0001-5002-0227 (2002) Microsatellite genotyping by primer extension and MALDI-ToF mass spectrometry. Plant Molecular Biology Reporter, 20 (3). pp. 259-263.

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A novel method for genotyping microsatellite alleles using primer extensions and mass spectrometry analysis has been developed. Following PCR amplification of the target region, a genotyping primer, with its 3′ end directly flanking the microsatellite repeats, was extended by a mixture of dNTPs complementary to the nucleotides composing the microsatellite. The length and molecular weight of extended primers vary with the number of repeats present in the allele(s) under examination. The weights of extension products were determined using matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-ToF MS) and used to identify genotypes on the basis of differential primer extension. This is a platform that is not gel based and is amenable to multiplexing and automation. The technique enables identification of heterozygous progeny in which alleles differ by a single trinucleotide repeat. The method is illustrated by genotyping a polymorphic microsatellite identified in an intron of the barley Mlo gene.

Item Type: Journal Article
Murdoch Affiliation(s): Western Australian State Agricultural Biotechnology Centre
Publisher: Springer New York
Copyright: © Springer 2002
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