Application of a double-enrichment procedure for microsatellite isolation and the use of tailed primers for high throughput genotyping
Autor(a) principal: | |
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Data de Publicação: | 2007 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Genetics and Molecular Biology |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572007000300014 |
Resumo: | 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. |
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Genetics and Molecular Biology |
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Application of a double-enrichment procedure for microsatellite isolation and the use of tailed primers for high throughput genotypingdouble-enrichmentdouble-hybridizationstutter bandstailed microsatellite primersThe 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.Sociedade Brasileira de Genética2007-03-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572007000300014Genetics and Molecular Biology v.30 n.2 2007reponame:Genetics and Molecular Biologyinstname:Sociedade Brasileira de Genética (SBG)instacron:SBG10.1590/S1415-47572007000300014info:eu-repo/semantics/openAccessDiniz,Fábio MendonçaIyengar,AratiLima,Paulo Sarmanho da CostaMaclean,NormanBentzen,Pauleng2007-06-04T00:00:00Zoai:scielo:S1415-47572007000300014Revistahttp://www.gmb.org.br/ONGhttps://old.scielo.br/oai/scielo-oai.php||editor@gmb.org.br1678-46851415-4757opendoar:2007-06-04T00:00Genetics and Molecular Biology - Sociedade Brasileira de Genética (SBG)false |
dc.title.none.fl_str_mv |
Application of a double-enrichment procedure for microsatellite isolation and the use of tailed primers for high throughput genotyping |
title |
Application of a double-enrichment procedure for microsatellite isolation and the use of tailed primers for high throughput genotyping |
spellingShingle |
Application of a double-enrichment procedure for microsatellite isolation and the use of tailed primers for high throughput genotyping Diniz,Fábio Mendonça double-enrichment double-hybridization stutter bands tailed microsatellite primers |
title_short |
Application of a double-enrichment procedure for microsatellite isolation and the use of tailed primers for high throughput genotyping |
title_full |
Application of a double-enrichment procedure for microsatellite isolation and the use of tailed primers for high throughput genotyping |
title_fullStr |
Application of a double-enrichment procedure for microsatellite isolation and the use of tailed primers for high throughput genotyping |
title_full_unstemmed |
Application of a double-enrichment procedure for microsatellite isolation and the use of tailed primers for high throughput genotyping |
title_sort |
Application of a double-enrichment procedure for microsatellite isolation and the use of tailed primers for high throughput genotyping |
author |
Diniz,Fábio Mendonça |
author_facet |
Diniz,Fábio Mendonça Iyengar,Arati Lima,Paulo Sarmanho da Costa Maclean,Norman Bentzen,Paul |
author_role |
author |
author2 |
Iyengar,Arati Lima,Paulo Sarmanho da Costa Maclean,Norman Bentzen,Paul |
author2_role |
author author author author |
dc.contributor.author.fl_str_mv |
Diniz,Fábio Mendonça Iyengar,Arati Lima,Paulo Sarmanho da Costa Maclean,Norman Bentzen,Paul |
dc.subject.por.fl_str_mv |
double-enrichment double-hybridization stutter bands tailed microsatellite primers |
topic |
double-enrichment double-hybridization stutter bands tailed microsatellite primers |
description |
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. |
publishDate |
2007 |
dc.date.none.fl_str_mv |
2007-03-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572007000300014 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572007000300014 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S1415-47572007000300014 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Sociedade Brasileira de Genética |
publisher.none.fl_str_mv |
Sociedade Brasileira de Genética |
dc.source.none.fl_str_mv |
Genetics and Molecular Biology v.30 n.2 2007 reponame:Genetics and Molecular Biology instname:Sociedade Brasileira de Genética (SBG) instacron:SBG |
instname_str |
Sociedade Brasileira de Genética (SBG) |
instacron_str |
SBG |
institution |
SBG |
reponame_str |
Genetics and Molecular Biology |
collection |
Genetics and Molecular Biology |
repository.name.fl_str_mv |
Genetics and Molecular Biology - Sociedade Brasileira de Genética (SBG) |
repository.mail.fl_str_mv |
||editor@gmb.org.br |
_version_ |
1752122380643205120 |