Influence of the electrokinetic injection conditions on the separation of DNA fragments in capillary electrophoresis
Autor(a) principal: | |
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Data de Publicação: | 2004 |
Outros Autores: | |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Journal of the Brazilian Chemical Society (Online) |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-50532004000300012 |
Resumo: | In genetic analysis by capillary electrophoresis with polymer solutions there are many variables that affect separation of the DNA fragments. A very critical one is the sample injection process, which can considerably affect the peak efficiency and the resolution. In this work, we have studied the influence of the DNA sample composition and the electrokinetic injection conditions in the separation of DNA fragments by capillary electrophoresis using replaceable polymer solutions. The studies were carried out by electrokinetically injecting the DNA samples under a variety of conditions, such as sample ionic strength, buffering capacity and the quality of the separation matrix. In all experiments DNA was stained with ethidium bromide for laser-induced fluorescence (LIF) detection and separated in a poly(vinylalcohol) coated capillary column filled with 0.5% hydroxyethylcellulose solution under a 200 V cm-1 electric field. Under such conditions, samples prepared with 1 to 5 mmol L-1 of running buffer have been shown to produce reproducible injections with RSD < 4% for both migration time and peak area. |
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Influence of the electrokinetic injection conditions on the separation of DNA fragments in capillary electrophoresisDNA sizingpolymer solutionselectrokinetic injectionlaser-induced fluorescenceIn genetic analysis by capillary electrophoresis with polymer solutions there are many variables that affect separation of the DNA fragments. A very critical one is the sample injection process, which can considerably affect the peak efficiency and the resolution. In this work, we have studied the influence of the DNA sample composition and the electrokinetic injection conditions in the separation of DNA fragments by capillary electrophoresis using replaceable polymer solutions. The studies were carried out by electrokinetically injecting the DNA samples under a variety of conditions, such as sample ionic strength, buffering capacity and the quality of the separation matrix. In all experiments DNA was stained with ethidium bromide for laser-induced fluorescence (LIF) detection and separated in a poly(vinylalcohol) coated capillary column filled with 0.5% hydroxyethylcellulose solution under a 200 V cm-1 electric field. Under such conditions, samples prepared with 1 to 5 mmol L-1 of running buffer have been shown to produce reproducible injections with RSD < 4% for both migration time and peak area.Sociedade Brasileira de Química2004-06-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-50532004000300012Journal of the Brazilian Chemical Society v.15 n.3 2004reponame:Journal of the Brazilian Chemical Society (Online)instname:Sociedade Brasileira de Química (SBQ)instacron:SBQ10.1590/S0103-50532004000300012info:eu-repo/semantics/openAccessCatai,Jonatan RicardoCarrilho,Emanueleng2004-06-16T00:00:00Zoai:scielo:S0103-50532004000300012Revistahttp://jbcs.sbq.org.brONGhttps://old.scielo.br/oai/scielo-oai.php||office@jbcs.sbq.org.br1678-47900103-5053opendoar:2004-06-16T00:00Journal of the Brazilian Chemical Society (Online) - Sociedade Brasileira de Química (SBQ)false |
dc.title.none.fl_str_mv |
Influence of the electrokinetic injection conditions on the separation of DNA fragments in capillary electrophoresis |
title |
Influence of the electrokinetic injection conditions on the separation of DNA fragments in capillary electrophoresis |
spellingShingle |
Influence of the electrokinetic injection conditions on the separation of DNA fragments in capillary electrophoresis Catai,Jonatan Ricardo DNA sizing polymer solutions electrokinetic injection laser-induced fluorescence |
title_short |
Influence of the electrokinetic injection conditions on the separation of DNA fragments in capillary electrophoresis |
title_full |
Influence of the electrokinetic injection conditions on the separation of DNA fragments in capillary electrophoresis |
title_fullStr |
Influence of the electrokinetic injection conditions on the separation of DNA fragments in capillary electrophoresis |
title_full_unstemmed |
Influence of the electrokinetic injection conditions on the separation of DNA fragments in capillary electrophoresis |
title_sort |
Influence of the electrokinetic injection conditions on the separation of DNA fragments in capillary electrophoresis |
author |
Catai,Jonatan Ricardo |
author_facet |
Catai,Jonatan Ricardo Carrilho,Emanuel |
author_role |
author |
author2 |
Carrilho,Emanuel |
author2_role |
author |
dc.contributor.author.fl_str_mv |
Catai,Jonatan Ricardo Carrilho,Emanuel |
dc.subject.por.fl_str_mv |
DNA sizing polymer solutions electrokinetic injection laser-induced fluorescence |
topic |
DNA sizing polymer solutions electrokinetic injection laser-induced fluorescence |
description |
In genetic analysis by capillary electrophoresis with polymer solutions there are many variables that affect separation of the DNA fragments. A very critical one is the sample injection process, which can considerably affect the peak efficiency and the resolution. In this work, we have studied the influence of the DNA sample composition and the electrokinetic injection conditions in the separation of DNA fragments by capillary electrophoresis using replaceable polymer solutions. The studies were carried out by electrokinetically injecting the DNA samples under a variety of conditions, such as sample ionic strength, buffering capacity and the quality of the separation matrix. In all experiments DNA was stained with ethidium bromide for laser-induced fluorescence (LIF) detection and separated in a poly(vinylalcohol) coated capillary column filled with 0.5% hydroxyethylcellulose solution under a 200 V cm-1 electric field. Under such conditions, samples prepared with 1 to 5 mmol L-1 of running buffer have been shown to produce reproducible injections with RSD < 4% for both migration time and peak area. |
publishDate |
2004 |
dc.date.none.fl_str_mv |
2004-06-01 |
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=S0103-50532004000300012 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-50532004000300012 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S0103-50532004000300012 |
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 Química |
publisher.none.fl_str_mv |
Sociedade Brasileira de Química |
dc.source.none.fl_str_mv |
Journal of the Brazilian Chemical Society v.15 n.3 2004 reponame:Journal of the Brazilian Chemical Society (Online) instname:Sociedade Brasileira de Química (SBQ) instacron:SBQ |
instname_str |
Sociedade Brasileira de Química (SBQ) |
instacron_str |
SBQ |
institution |
SBQ |
reponame_str |
Journal of the Brazilian Chemical Society (Online) |
collection |
Journal of the Brazilian Chemical Society (Online) |
repository.name.fl_str_mv |
Journal of the Brazilian Chemical Society (Online) - Sociedade Brasileira de Química (SBQ) |
repository.mail.fl_str_mv |
||office@jbcs.sbq.org.br |
_version_ |
1750318165746778112 |