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

Autores
Ginart, Santiago; Caputo, Mariela; Corach, Daniel; Sala, Adriana Andrea
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
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.
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; Argentina
Fil: Caputo, Mariela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina
Fil: Corach, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina
Fil: 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; Argentina
Materia
Degraded Dna Assessment
Dna Quantitation
High Resolution Melting Analysis
Male Dna Detection
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/48625
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/48625
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
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 Técnicas2025-12-03T08:33:52Zoai: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:34982025-12-03 08:33:52.955CONICET 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
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
topic Degraded Dna Assessment
Dna Quantitation
High Resolution Melting Analysis
Male Dna Detection
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv 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.
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; Argentina
Fil: Caputo, Mariela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina
Fil: Corach, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina
Fil: 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; Argentina
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
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
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 13.214268

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