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Registro Completo |
Biblioteca(s): |
Embrapa Meio-Norte. |
Data corrente: |
10/03/2008 |
Data da última atualização: |
14/09/2023 |
Tipo da produção científica: |
Artigo em Periódico Indexado |
Autoria: |
DINIZ, F. M.; IYENGAR, A.; LIMA, P. S. da C.; MACLEAN, N.; BENTZEN, P. |
Afiliação: |
FABIO MENDONCA DINIZ, CPAMN; ARATI IYENGAR; PAULO SARMANHO DA COSTA LIMA, CPAMN; NORMAN MACLEAN; PAUL BENTZEN. |
Título: |
Application of a double-enrichment procedure for microsatellite isolation and the use tailed primers for high throughput genotyping. |
Ano de publicação: |
2007 |
Fonte/Imprenta: |
Genetics and Molecular Biology, São Paulo, v. 30, n. 2, p. 380-384, mar. 2007. |
ISSN: |
1415-4757 |
Idioma: |
Inglês |
Conteúdo: |
The number of microsatellite loci and their allelic diversity contribute to increase accuracy and informativity of genetic estimates, however, the isolation of microsatellite loci is not only laborious but also quite expensive. We used (GATA)n and (GACA)n tetranucleotide probes and single- and double-enrichment hybridization to construct and screen a genomic library with an increased proportion of DNA fragments containing repeat motifs. Repeats were found using both types of hybridization but the double-enrichment procedure recovered sequences of which 100% contained (GATA)n and (GACA)n motifs. Microsatellite loci primers were then designed with an M13R-tail or CAG-tag to produce scorable PCR products with minimal stutter. The approach used in this study suggests that double-enrichment is a worthwhile strategy when isolating repeat motifs from eukaryotic genomes. Moreover, the use of tailed microsatellite primers provides increased resolution for compound microsatellite loci, with a significant decrease in costs. |
Palavras-Chave: |
Banda polimórfica; double-enrichment; double-hybridization; Enriquecimento duplo; Hibridação dupla; Microssatélite; stutter bands; tailed microsatellite primer. |
Categoria do assunto: |
X Pesquisa, Tecnologia e Engenharia |
URL: |
https://ainfo.cnptia.embrapa.br/digital/bitstream/doc/65872/1/Application-21449.pdf
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Marc: |
LEADER 01914naa a2200277 a 4500 001 1065872 005 2023-09-14 008 2007 bl uuuu u00u1 u #d 022 $a1415-4757 100 1 $aDINIZ, F. M. 245 $aApplication of a double-enrichment procedure for microsatellite isolation and the use tailed primers for high throughput genotyping.$h[electronic resource] 260 $c2007 520 $aThe number of microsatellite loci and their allelic diversity contribute to increase accuracy and informativity of genetic estimates, however, the isolation of microsatellite loci is not only laborious but also quite expensive. We used (GATA)n and (GACA)n tetranucleotide probes and single- and double-enrichment hybridization to construct and screen a genomic library with an increased proportion of DNA fragments containing repeat motifs. Repeats were found using both types of hybridization but the double-enrichment procedure recovered sequences of which 100% contained (GATA)n and (GACA)n motifs. Microsatellite loci primers were then designed with an M13R-tail or CAG-tag to produce scorable PCR products with minimal stutter. The approach used in this study suggests that double-enrichment is a worthwhile strategy when isolating repeat motifs from eukaryotic genomes. Moreover, the use of tailed microsatellite primers provides increased resolution for compound microsatellite loci, with a significant decrease in costs. 653 $aBanda polimórfica 653 $adouble-enrichment 653 $adouble-hybridization 653 $aEnriquecimento duplo 653 $aHibridação dupla 653 $aMicrossatélite 653 $astutter bands 653 $atailed microsatellite primer 700 1 $aIYENGAR, A. 700 1 $aLIMA, P. S. da C. 700 1 $aMACLEAN, N. 700 1 $aBENTZEN, P. 773 $tGenetics and Molecular Biology, São Paulo$gv. 30, n. 2, p. 380-384, mar. 2007.
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