PCR-mediated recombination in development of microsatellite markers: mechanism and implications



Título del documento: PCR-mediated recombination in development of microsatellite markers: mechanism and implications
Revista: Genetics and molecular biology
Base de datos: PERIÓDICA
Número de sistema: 000331325
ISSN: 1415-4757
Autores: 1
1
1
1
Instituciones: 1Universidade Federal de Santa Maria, Centro de Ciencias Naturais e Exatas, Santa Maria, Rio Grande do Sul. Brasil
Año:
Periodo: Mar
Volumen: 31
Número: 1
Paginación: 58=63-58=63
País: Brasil
Idioma: Inglés
Tipo de documento: Artículo
Enfoque: Experimental
Resumen en inglés 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
Disciplinas: Biología,
Química
Palabras clave: Crustáceos,
Genética,
Bioquímica,
Producto quimerico de PCR,
Aislamiento de microsatélites,
Recombinación,
Secuencia simple,
Aegla longirostri
Keyword: Biology,
Chemistry,
Crustaceans,
Genetics,
Biochemistry,
Chimeric PCR product,
Microsatellite isolation,
Recombination,
Simple sequence,
Aegla longirostri
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