A simple and cost-effective protocol for DNA isolation from buccal epithelial cells

Detalhes bibliográficos
Autor(a) principal: Aidar,Marisi
Data de Publicação: 2007
Outros Autores: Line,Sergio Roberto Peres
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Brazilian Dental Journal
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-64402007000200012
Resumo: Buccal cells provide a convenient source of DNA for epidemiological studies. The goal of this study was to develop a convenient method to obtain buccal cells from mouthwash samples to be used as a source of DNA, and to evaluate the stability of the DNA in mouthwash solution over time. The procedures used in the method described in this paper avoid the use of any organic solvents. This is achieved by salting out the cellular proteins by dehydration and precipitation with a saturated ammonium acetate solution. The protocol described here is fast, simple to perform, sensitive, economical and several samples can be processed at the same time. The analyses provide consistent evidence that DNA extracted by this methodology is sufficient for several PCR amplifications. The total DNA yield ranged from 5 to 93 µg (median 15 µg, mean 20.71 µg). DNA can be extracted and PCR amplified after storage of mouthwash solution at room temperature for periods of up to 30 days.
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spelling A simple and cost-effective protocol for DNA isolation from buccal epithelial cellsbuccal cellsdna extractionammonium acetateBuccal cells provide a convenient source of DNA for epidemiological studies. The goal of this study was to develop a convenient method to obtain buccal cells from mouthwash samples to be used as a source of DNA, and to evaluate the stability of the DNA in mouthwash solution over time. The procedures used in the method described in this paper avoid the use of any organic solvents. This is achieved by salting out the cellular proteins by dehydration and precipitation with a saturated ammonium acetate solution. The protocol described here is fast, simple to perform, sensitive, economical and several samples can be processed at the same time. The analyses provide consistent evidence that DNA extracted by this methodology is sufficient for several PCR amplifications. The total DNA yield ranged from 5 to 93 µg (median 15 µg, mean 20.71 µg). DNA can be extracted and PCR amplified after storage of mouthwash solution at room temperature for periods of up to 30 days.Fundação Odontológica de Ribeirão Preto2007-01-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-64402007000200012Brazilian Dental Journal v.18 n.2 2007reponame:Brazilian Dental Journalinstname:Fundação Odontológica de Ribeirão Preto (FUNORP)instacron:FUNORP10.1590/S0103-64402007000200012info:eu-repo/semantics/openAccessAidar,MarisiLine,Sergio Roberto Pereseng2007-10-25T00:00:00Zoai:scielo:S0103-64402007000200012Revistahttps://www.scielo.br/j/bdj/https://old.scielo.br/oai/scielo-oai.phpbdj@forp.usp.br||sergio@fosjc.unesp.br1806-47600103-6440opendoar:2007-10-25T00:00Brazilian Dental Journal - Fundação Odontológica de Ribeirão Preto (FUNORP)false
dc.title.none.fl_str_mv A simple and cost-effective protocol for DNA isolation from buccal epithelial cells
title A simple and cost-effective protocol for DNA isolation from buccal epithelial cells
spellingShingle A simple and cost-effective protocol for DNA isolation from buccal epithelial cells
Aidar,Marisi
buccal cells
dna extraction
ammonium acetate
title_short A simple and cost-effective protocol for DNA isolation from buccal epithelial cells
title_full A simple and cost-effective protocol for DNA isolation from buccal epithelial cells
title_fullStr A simple and cost-effective protocol for DNA isolation from buccal epithelial cells
title_full_unstemmed A simple and cost-effective protocol for DNA isolation from buccal epithelial cells
title_sort A simple and cost-effective protocol for DNA isolation from buccal epithelial cells
author Aidar,Marisi
author_facet Aidar,Marisi
Line,Sergio Roberto Peres
author_role author
author2 Line,Sergio Roberto Peres
author2_role author
dc.contributor.author.fl_str_mv Aidar,Marisi
Line,Sergio Roberto Peres
dc.subject.por.fl_str_mv buccal cells
dna extraction
ammonium acetate
topic buccal cells
dna extraction
ammonium acetate
description Buccal cells provide a convenient source of DNA for epidemiological studies. The goal of this study was to develop a convenient method to obtain buccal cells from mouthwash samples to be used as a source of DNA, and to evaluate the stability of the DNA in mouthwash solution over time. The procedures used in the method described in this paper avoid the use of any organic solvents. This is achieved by salting out the cellular proteins by dehydration and precipitation with a saturated ammonium acetate solution. The protocol described here is fast, simple to perform, sensitive, economical and several samples can be processed at the same time. The analyses provide consistent evidence that DNA extracted by this methodology is sufficient for several PCR amplifications. The total DNA yield ranged from 5 to 93 µg (median 15 µg, mean 20.71 µg). DNA can be extracted and PCR amplified after storage of mouthwash solution at room temperature for periods of up to 30 days.
publishDate 2007
dc.date.none.fl_str_mv 2007-01-01
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=S0103-64402007000200012
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-64402007000200012
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/S0103-64402007000200012
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 Fundação Odontológica de Ribeirão Preto
publisher.none.fl_str_mv Fundação Odontológica de Ribeirão Preto
dc.source.none.fl_str_mv Brazilian Dental Journal v.18 n.2 2007
reponame:Brazilian Dental Journal
instname:Fundação Odontológica de Ribeirão Preto (FUNORP)
instacron:FUNORP
instname_str Fundação Odontológica de Ribeirão Preto (FUNORP)
instacron_str FUNORP
institution FUNORP
reponame_str Brazilian Dental Journal
collection Brazilian Dental Journal
repository.name.fl_str_mv Brazilian Dental Journal - Fundação Odontológica de Ribeirão Preto (FUNORP)
repository.mail.fl_str_mv bdj@forp.usp.br||sergio@fosjc.unesp.br
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