Journal article
Microsatellite genotyping by primer extension and MALDI-ToF mass spectrometry
Plant Molecular Biology Reporter, Vol.20(3), pp.259-263
2002
Abstract
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.
Details
- Title
- Microsatellite genotyping by primer extension and MALDI-ToF mass spectrometry
- Authors/Creators
- M. Paris (Author/Creator) - Murdoch UniversityM.G.K. Jones (Author/Creator) - Murdoch University
- Publication Details
- Plant Molecular Biology Reporter, Vol.20(3), pp.259-263
- Publisher
- Springer New York
- Identifiers
- 991005541227307891
- Copyright
- © Springer 2002
- Murdoch Affiliation
- State Agricultural Biotechnology Centre
- Language
- English
- Resource Type
- Journal article
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- Citation topics
- 3 Agriculture, Environment & Ecology
- 3.4 Crop Science
- 3.4.96 QTL
- Web Of Science research areas
- Biochemical Research Methods
- Plant Sciences
- ESI research areas
- Plant & Animal Science