Measuring human DNA degradation and gender detection in forensic DNA samples by q-PCR/HRM analysis

Human DNA quantification, DNA degradation assessment and gender determination are key aspects in most field of human DNA analysis. The assay reported here is a tri-plex Real Time quantitative PCR reaction followed by high resolution melting (HRM) using Syto9 as intercalating dye. The system produces...

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Detalhes bibliográficos
Autores: Ginart, Santiago, Caputo, Mariela, Corach, Daniel, Sala, Adriana Andrea
Tipo de documento: artigo
Estado:Versão publicada
Data de publicação:2017
País:Argentina
Recursos:Consejo Nacional de Investigaciones Científicas y Técnicas
Repositório:CONICET Digital (CONICET)
Idioma:inglês
OAI Identifier:oai:ri.conicet.gov.ar:11336/48625
Acesso em linha:http://hdl.handle.net/11336/48625
Access Level:Acceso aberto
Palavra-chave:Degraded Dna Assessment
Dna Quantitation
High Resolution Melting Analysis
Male Dna Detection
https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
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spelling Measuring human DNA degradation and gender detection in forensic DNA samples by q-PCR/HRM analysisGinart, SantiagoCaputo, MarielaCorach, DanielSala, Adriana AndreaDegraded Dna AssessmentDna QuantitationHigh Resolution Melting AnalysisMale Dna Detectionhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Human DNA quantification, DNA degradation assessment and gender determination are key aspects in most field of human DNA analysis. The assay reported here is a tri-plex Real Time quantitative PCR reaction followed by high resolution melting (HRM) using Syto9 as intercalating dye. The system produces three amplicons: 1- transducin (beta)-like 1, Y-linked −TBL1Y (84 bp), 2- DeGraded small target DNA–DGst- (152 bp) and 3-DeGraded large target DNA-DGlt- (244 bp). DNA quantitation is based on total fluorescence; TBL1Y amplicon allows detecting male DNA and the ratio DGst/DGlt to assess DNA degradation level. q-PCR quantitation proved good linearity in triplicates among 3.2 pg/ul–50 ng/ul DNA concentration range. Amplification efficiency (E) and reaction slope (m) mean values were 1.04 and 3.23 respectively. Upon HRM analysis, three melting peaks are detected in a male DNA sample and two if only female DNA is present. We define the parameter D as the ratio DGst/DGlt that reflects the extent of DNA degradation in a given sample. A direct correlation has been demonstrated between DNA damage and increased value of parameter D. This q-PCR approach is rapid, sensitive, and a cost-effective method suitable for detecting degraded DNA samples and applicable to any field where human DNA quantitation-qualification is required.Fil: Ginart, Santiago. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; ArgentinaFil: Caputo, Mariela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; ArgentinaFil: Corach, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; ArgentinaFil: Sala, Adriana Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; ArgentinaElsevier Science2017-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/48625Ginart, Santiago; Caputo, Mariela; Corach, Daniel; Sala, Adriana Andrea; Measuring human DNA degradation and gender detection in forensic DNA samples by q-PCR/HRM analysis; Elsevier Science; Forensic Science International: Genetics Supplement Series; 6; 12-2017; e552-e5541875-175XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.fsigss.2017.09.211info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S1875176817301038info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2024-05-08T13:32:58Zoai:ri.conicet.gov.ar:11336/48625instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982024-05-08 13:32:58.926CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Measuring human DNA degradation and gender detection in forensic DNA samples by q-PCR/HRM analysis
title Measuring human DNA degradation and gender detection in forensic DNA samples by q-PCR/HRM analysis
spellingShingle Measuring human DNA degradation and gender detection in forensic DNA samples by q-PCR/HRM analysis
Ginart, Santiago
Degraded Dna Assessment
Dna Quantitation
High Resolution Melting Analysis
Male Dna Detection
https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
title_short Measuring human DNA degradation and gender detection in forensic DNA samples by q-PCR/HRM analysis
title_full Measuring human DNA degradation and gender detection in forensic DNA samples by q-PCR/HRM analysis
title_fullStr Measuring human DNA degradation and gender detection in forensic DNA samples by q-PCR/HRM analysis
title_full_unstemmed Measuring human DNA degradation and gender detection in forensic DNA samples by q-PCR/HRM analysis
title_sort Measuring human DNA degradation and gender detection in forensic DNA samples by q-PCR/HRM analysis
dc.creator.none.fl_str_mv Ginart, Santiago
Caputo, Mariela
Corach, Daniel
Sala, Adriana Andrea
author Ginart, Santiago
author_facet Ginart, Santiago
Caputo, Mariela
Corach, Daniel
Sala, Adriana Andrea
author_role author
author2 Caputo, Mariela
Corach, Daniel
Sala, Adriana Andrea
author2_role author
author
author
dc.subject.none.fl_str_mv Degraded Dna Assessment
Dna Quantitation
High Resolution Melting Analysis
Male Dna Detection
https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
topic Degraded Dna Assessment
Dna Quantitation
High Resolution Melting Analysis
Male Dna Detection
https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
description Human DNA quantification, DNA degradation assessment and gender determination are key aspects in most field of human DNA analysis. The assay reported here is a tri-plex Real Time quantitative PCR reaction followed by high resolution melting (HRM) using Syto9 as intercalating dye. The system produces three amplicons: 1- transducin (beta)-like 1, Y-linked −TBL1Y (84 bp), 2- DeGraded small target DNA–DGst- (152 bp) and 3-DeGraded large target DNA-DGlt- (244 bp). DNA quantitation is based on total fluorescence; TBL1Y amplicon allows detecting male DNA and the ratio DGst/DGlt to assess DNA degradation level. q-PCR quantitation proved good linearity in triplicates among 3.2 pg/ul–50 ng/ul DNA concentration range. Amplification efficiency (E) and reaction slope (m) mean values were 1.04 and 3.23 respectively. Upon HRM analysis, three melting peaks are detected in a male DNA sample and two if only female DNA is present. We define the parameter D as the ratio DGst/DGlt that reflects the extent of DNA degradation in a given sample. A direct correlation has been demonstrated between DNA damage and increased value of parameter D. This q-PCR approach is rapid, sensitive, and a cost-effective method suitable for detecting degraded DNA samples and applicable to any field where human DNA quantitation-qualification is required.
publishDate 2017
dc.date.none.fl_str_mv 2017-12
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/48625
Ginart, Santiago; Caputo, Mariela; Corach, Daniel; Sala, Adriana Andrea; Measuring human DNA degradation and gender detection in forensic DNA samples by q-PCR/HRM analysis; Elsevier Science; Forensic Science International: Genetics Supplement Series; 6; 12-2017; e552-e554
1875-175X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/48625
identifier_str_mv Ginart, Santiago; Caputo, Mariela; Corach, Daniel; Sala, Adriana Andrea; Measuring human DNA degradation and gender detection in forensic DNA samples by q-PCR/HRM analysis; Elsevier Science; Forensic Science International: Genetics Supplement Series; 6; 12-2017; e552-e554
1875-175X
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.fsigss.2017.09.211
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S1875176817301038
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Science
publisher.none.fl_str_mv Elsevier Science
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 15,228081

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