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Provedor de dados:  Genet. Mol. Biol.
País:  Brazil
Título:  PCR-mediated recombination in development of microsatellite markers: mechanism and implications
Autores:  Roratto,Paula A.
Buchmann,Darine
Santos,Sandro
Bartholomei-Santos,Marlise L.
Data:  2008-01-01
Ano:  2008
Palavras-chave:  Chimeric PCR product
Microsatellite isolation
Recombination
Simple sequence repeats (SSR)
Resumo:  Protocols for microsatellite-enrichment libraries have been widely applied to several species in order to supply the most informative molecular markers for population and inbreeding studies. One drawback of these protocols is the ratio of designed primer pairs that fail to amplify the expected fragment, even after exhaustive optimization attempts. A possible cause of unsuccessful microsatellite primers may be that such loci are artifacts resulting from chimeric PCR products, instead of real genomic sequences. The microsatellite-enriched library constructed for Aegla longirostri (Crustacea, Decapoda, Anomura) showed that 29% of sequenced clones were chimeric products because these sequences shared one of the flanking regions around the same repeat motif but not the other. PCR-mediated recombination is a well-known event described for several procedures in which related sequences are used as a template. We have associated this phenomenon with microsatellite marker development. This study explained the high ratio of recombinant sequences generated in the A. longirostri microsatellite-enriched library. We discuss the mechanism and implications of PCR chimeric-product formation during microsatellite isolation.
Tipo:  Info:eu-repo/semantics/other
Idioma:  Inglês
Identificador:  http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572008000100012
Editor:  Sociedade Brasileira de Genética
Relação:  10.1590/S1415-47572008000100012
Formato:  text/html
Fonte:  Genetics and Molecular Biology v.31 n.1 2008
Direitos:  info:eu-repo/semantics/openAccess
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