Optimization of PCR amplification of maize microsatellite loci
Autor(a) principal: | |
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Data de Publicação: | 2000 |
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=S1415-47572000000200026 |
Resumo: | Maize (Zea mays L.) microsatellite loci are useful as genetic markers because they are numerous, occur in every chromosome, and have a high content of polymorphism information. Furthermore, they can be amplified by PCR and the resulting fragments resolved on agarose gels. A major problem, however, is that the primers used in their amplification often have different length and nucleic acid content, requiring optimization of PCR amplification programs for each locus. We developed "touchdown" PCR programs successfully used in the amplification of a set of 125-maize microsatellite loci, chosen as representative of all chromosomes. The programs varied both in relation to the annealing temperature (Tm) and primer concentration. Primers could be divided into four groups. A large group included primers that amplified well in a basic previously published amplification program but with different primer concentrations. A second group amplified in alternative programs in which the annealing temperatures were changed so as to include the Tm of either both primers or the highest Tm of the primer pair estimated based on their nucleotide composition. A third group included primers that amplified in programs with Tm higher than those estimated, and in a fourth group, with just two pairs of primers, the opposite situation prevailed. |
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Genetics and Molecular Biology |
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Optimization of PCR amplification of maize microsatellite lociMaize (Zea mays L.) microsatellite loci are useful as genetic markers because they are numerous, occur in every chromosome, and have a high content of polymorphism information. Furthermore, they can be amplified by PCR and the resulting fragments resolved on agarose gels. A major problem, however, is that the primers used in their amplification often have different length and nucleic acid content, requiring optimization of PCR amplification programs for each locus. We developed "touchdown" PCR programs successfully used in the amplification of a set of 125-maize microsatellite loci, chosen as representative of all chromosomes. The programs varied both in relation to the annealing temperature (Tm) and primer concentration. Primers could be divided into four groups. A large group included primers that amplified well in a basic previously published amplification program but with different primer concentrations. A second group amplified in alternative programs in which the annealing temperatures were changed so as to include the Tm of either both primers or the highest Tm of the primer pair estimated based on their nucleotide composition. A third group included primers that amplified in programs with Tm higher than those estimated, and in a fourth group, with just two pairs of primers, the opposite situation prevailed.Sociedade Brasileira de Genética2000-06-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572000000200026Genetics and Molecular Biology v.23 n.2 2000reponame:Genetics and Molecular Biologyinstname:Sociedade Brasileira de Genética (SBG)instacron:SBG10.1590/S1415-47572000000200026info:eu-repo/semantics/openAccessOgliari,Juliana BernardiBoscariol,Raquel L.Camargo,Luis E.A.eng2000-09-22T00:00:00Zoai:scielo:S1415-47572000000200026Revistahttp://www.gmb.org.br/ONGhttps://old.scielo.br/oai/scielo-oai.php||editor@gmb.org.br1678-46851415-4757opendoar:2000-09-22T00:00Genetics and Molecular Biology - Sociedade Brasileira de Genética (SBG)false |
dc.title.none.fl_str_mv |
Optimization of PCR amplification of maize microsatellite loci |
title |
Optimization of PCR amplification of maize microsatellite loci |
spellingShingle |
Optimization of PCR amplification of maize microsatellite loci Ogliari,Juliana Bernardi |
title_short |
Optimization of PCR amplification of maize microsatellite loci |
title_full |
Optimization of PCR amplification of maize microsatellite loci |
title_fullStr |
Optimization of PCR amplification of maize microsatellite loci |
title_full_unstemmed |
Optimization of PCR amplification of maize microsatellite loci |
title_sort |
Optimization of PCR amplification of maize microsatellite loci |
author |
Ogliari,Juliana Bernardi |
author_facet |
Ogliari,Juliana Bernardi Boscariol,Raquel L. Camargo,Luis E.A. |
author_role |
author |
author2 |
Boscariol,Raquel L. Camargo,Luis E.A. |
author2_role |
author author |
dc.contributor.author.fl_str_mv |
Ogliari,Juliana Bernardi Boscariol,Raquel L. Camargo,Luis E.A. |
description |
Maize (Zea mays L.) microsatellite loci are useful as genetic markers because they are numerous, occur in every chromosome, and have a high content of polymorphism information. Furthermore, they can be amplified by PCR and the resulting fragments resolved on agarose gels. A major problem, however, is that the primers used in their amplification often have different length and nucleic acid content, requiring optimization of PCR amplification programs for each locus. We developed "touchdown" PCR programs successfully used in the amplification of a set of 125-maize microsatellite loci, chosen as representative of all chromosomes. The programs varied both in relation to the annealing temperature (Tm) and primer concentration. Primers could be divided into four groups. A large group included primers that amplified well in a basic previously published amplification program but with different primer concentrations. A second group amplified in alternative programs in which the annealing temperatures were changed so as to include the Tm of either both primers or the highest Tm of the primer pair estimated based on their nucleotide composition. A third group included primers that amplified in programs with Tm higher than those estimated, and in a fourth group, with just two pairs of primers, the opposite situation prevailed. |
publishDate |
2000 |
dc.date.none.fl_str_mv |
2000-06-01 |
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=S1415-47572000000200026 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572000000200026 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S1415-47572000000200026 |
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.23 n.2 2000 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_ |
1752122377752281088 |