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Development of a cost-effective pyrosequencing approach for SNP genotyping in barley

AuthorsSilvar Casao, Cristina CSIC ORCID; Perovic, Dragan; Casas Cendoya, Ana María CSIC ORCID ; Igartua Arregui, Ernesto CSIC ORCID ; Ordon, Frank
M13 tail
marker-assisted selection
Issue DateJun-2011
PublisherJohn Wiley & Sons
CitationSilvar C, Perovic D, Casas AM, Igartua E, Ordon F. Development of a cost-effective pyrosequencing approach for SNP genotyping in barley. Plant breeding 130 (3): 394-397 (2011)
AbstractAn improved, efficient, reliable and cost-effective pyrosequencing protocol for single nucleotide polymorphism (SNP) genotyping in plants is described. Labelling of the PCR products, required for the single-stranded pyrosequencing assay, is carried out in a one-step PCR with a universal biotinalyted M13 primer of 19-bp. A ratio of 1 : 10 of tailed primer to M13 primer in a 15-μl reaction volume turned out to be best suited for a successful identification of different alleles based on known SNPs. This technology allows cost-effective SNP genotyping in plant genomes for loci of interest at an overall low cost in a short period of time and is therefore best suited for marker-based selection procedures.
Description12 Pag., 1 Tabl., 1 Fig. The definitive version is available at: http://onlinelibrary.wiley.com/journal/10.1111/(ISSN)1439-0523
Publisher version (URL)http://dx.doi.org/10.1111/j.1439-0523.2011.01853.x
Appears in Collections:(EEAD) Artículos
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