Highly efficient automated extraction of DNA from old and contemporary skeletal remains
Abstract We optimised the automated extraction of DNA from old and contemporary skeletal remains using the AutoMate Express system and the PrepFiler BTA kit. 24 Contemporary and 25 old skeletal remains from WWII were analysed. For each skeleton, extraction using only 0.05 g of powder was performed a...
Gespeichert in:
Veröffentlicht in: | Journal of forensic and legal medicine 2016-01, Vol.37, p.78-86 |
---|---|
Hauptverfasser: | , , , , , , , |
Format: | Artikel |
Sprache: | eng |
Schlagworte: | |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
container_end_page | 86 |
---|---|
container_issue | |
container_start_page | 78 |
container_title | Journal of forensic and legal medicine |
container_volume | 37 |
creator | Zupanič Pajnič, Irena Debska, Magdalena Gornjak Pogorelc, Barbara Vodopivec Mohorčič, Katja Balažic, Jože Zupanc, Tomaž Štefanič, Borut Geršak, Ksenija |
description | Abstract We optimised the automated extraction of DNA from old and contemporary skeletal remains using the AutoMate Express system and the PrepFiler BTA kit. 24 Contemporary and 25 old skeletal remains from WWII were analysed. For each skeleton, extraction using only 0.05 g of powder was performed according to the manufacturer's recommendations (no demineralisation – ND method). Since only 32% of full profiles were obtained from aged and 58% from contemporary casework skeletons, the extraction protocol was modified to acquire higher quality DNA and genomic DNA was obtained after full demineralisation (FD method). The nuclear DNA of the samples was quantified using the Investigator Quantiplex kit and STR typing was performed using the NGM kit to evaluate the performance of tested extraction methods. In the aged DNA samples, 64% of full profiles were obtained using the FD method. For the contemporary skeletal remains the performance of the ND method was closer to the FD method compared to the old skeletons, giving 58% of full profiles with the ND method and 71% of full profiles using the FD method. The extraction of DNA from only 0.05 g of bone or tooth powder using the AutoMate Express has proven highly successful in the recovery of DNA from old and contemporary skeletons, especially with the modified FD method. We believe that the results obtained will contribute to the possibilities of using automated devices for extracting DNA from skeletal remains, which would shorten the procedures for obtaining high-quality DNA from skeletons in forensic laboratories. |
doi_str_mv | 10.1016/j.jflm.2015.11.001 |
format | Article |
fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_1751993394</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S1752928X15002073</els_id><sourcerecordid>3935299351</sourcerecordid><originalsourceid>FETCH-LOGICAL-c439t-bd03d7daad99e833db3b0576c14c46be9f6a4411a5bb73c42f2060b863ab106a3</originalsourceid><addsrcrecordid>eNp9kU1rFTEUhgdRbK3-ARcScONmpvmaZAZEKPWjQqkLFd2FTHJGM81MrkmmeP-9GW5V6KKrnMXzvuQ8p6qeE9wQTMTp1EyjnxuKSdsQ0mBMHlTHpJNdLXknH5ZZtrTuaff9qHqS0oRxy6lsH1dHVAjScsmPq28X7sdPv0cwjs44WDLSaw6zzmAR_M5Rm-zCgsKI3l6doTGGGQVvkV4sMmHJMO9C1HGP0jV4yNqjCLN2S3paPRq1T_Ds9j2pvr5_9-X8or789OHj-dllbTjrcz1YzKy0Wtu-h44xO7ABt1IYwg0XA_Sj0JwTotthkMxwOlIs8NAJpgeChWYn1atD7y6GXyukrGaXDHivFwhrUkUB6XvGel7Ql3fQKaxxKb8rlMAdL0JkoeiBMjGkFGFUu-jmsqIiWG3a1aQ27WrTrghRRXsJvbitXocZ7L_IX88FeH0AoLi4cRBV2mwbsC6CycoGd3__mztx493ijPbXsIf0fw-VqMLq83b47e6kxZhiydgf5oOoYQ</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1760844747</pqid></control><display><type>article</type><title>Highly efficient automated extraction of DNA from old and contemporary skeletal remains</title><source>MEDLINE</source><source>Access via ScienceDirect (Elsevier)</source><creator>Zupanič Pajnič, Irena ; Debska, Magdalena ; Gornjak Pogorelc, Barbara ; Vodopivec Mohorčič, Katja ; Balažic, Jože ; Zupanc, Tomaž ; Štefanič, Borut ; Geršak, Ksenija</creator><creatorcontrib>Zupanič Pajnič, Irena ; Debska, Magdalena ; Gornjak Pogorelc, Barbara ; Vodopivec Mohorčič, Katja ; Balažic, Jože ; Zupanc, Tomaž ; Štefanič, Borut ; Geršak, Ksenija</creatorcontrib><description>Abstract We optimised the automated extraction of DNA from old and contemporary skeletal remains using the AutoMate Express system and the PrepFiler BTA kit. 24 Contemporary and 25 old skeletal remains from WWII were analysed. For each skeleton, extraction using only 0.05 g of powder was performed according to the manufacturer's recommendations (no demineralisation – ND method). Since only 32% of full profiles were obtained from aged and 58% from contemporary casework skeletons, the extraction protocol was modified to acquire higher quality DNA and genomic DNA was obtained after full demineralisation (FD method). The nuclear DNA of the samples was quantified using the Investigator Quantiplex kit and STR typing was performed using the NGM kit to evaluate the performance of tested extraction methods. In the aged DNA samples, 64% of full profiles were obtained using the FD method. For the contemporary skeletal remains the performance of the ND method was closer to the FD method compared to the old skeletons, giving 58% of full profiles with the ND method and 71% of full profiles using the FD method. The extraction of DNA from only 0.05 g of bone or tooth powder using the AutoMate Express has proven highly successful in the recovery of DNA from old and contemporary skeletons, especially with the modified FD method. We believe that the results obtained will contribute to the possibilities of using automated devices for extracting DNA from skeletal remains, which would shorten the procedures for obtaining high-quality DNA from skeletons in forensic laboratories.</description><identifier>ISSN: 1752-928X</identifier><identifier>EISSN: 1878-7487</identifier><identifier>DOI: 10.1016/j.jflm.2015.11.001</identifier><identifier>PMID: 26615474</identifier><language>eng</language><publisher>England: Elsevier Ltd</publisher><subject>Bone and Bones - chemistry ; Bones ; Deoxyribonucleic acid ; DNA ; DNA - isolation & purification ; DNA extraction ; DNA Fingerprinting ; Efficiency ; Forensic Genetics - methods ; Forensic sciences ; History, 20th Century ; Human remains ; Humans ; Microsatellite Repeats ; Missing person identification ; Pathology ; Real-Time Polymerase Chain Reaction ; Second World War ; Skeletal remains ; Skeletal system ; Teeth ; Tooth - chemistry</subject><ispartof>Journal of forensic and legal medicine, 2016-01, Vol.37, p.78-86</ispartof><rights>Elsevier Ltd and Faculty of Forensic and Legal Medicine</rights><rights>2015 Elsevier Ltd and Faculty of Forensic and Legal Medicine</rights><rights>Copyright © 2015 Elsevier Ltd and Faculty of Forensic and Legal Medicine. All rights reserved.</rights><rights>Copyright Churchill Livingstone Inc., Medical Publishers Jan 2016</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c439t-bd03d7daad99e833db3b0576c14c46be9f6a4411a5bb73c42f2060b863ab106a3</citedby><cites>FETCH-LOGICAL-c439t-bd03d7daad99e833db3b0576c14c46be9f6a4411a5bb73c42f2060b863ab106a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.jflm.2015.11.001$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>315,781,785,3551,27928,27929,45999</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26615474$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zupanič Pajnič, Irena</creatorcontrib><creatorcontrib>Debska, Magdalena</creatorcontrib><creatorcontrib>Gornjak Pogorelc, Barbara</creatorcontrib><creatorcontrib>Vodopivec Mohorčič, Katja</creatorcontrib><creatorcontrib>Balažic, Jože</creatorcontrib><creatorcontrib>Zupanc, Tomaž</creatorcontrib><creatorcontrib>Štefanič, Borut</creatorcontrib><creatorcontrib>Geršak, Ksenija</creatorcontrib><title>Highly efficient automated extraction of DNA from old and contemporary skeletal remains</title><title>Journal of forensic and legal medicine</title><addtitle>J Forensic Leg Med</addtitle><description>Abstract We optimised the automated extraction of DNA from old and contemporary skeletal remains using the AutoMate Express system and the PrepFiler BTA kit. 24 Contemporary and 25 old skeletal remains from WWII were analysed. For each skeleton, extraction using only 0.05 g of powder was performed according to the manufacturer's recommendations (no demineralisation – ND method). Since only 32% of full profiles were obtained from aged and 58% from contemporary casework skeletons, the extraction protocol was modified to acquire higher quality DNA and genomic DNA was obtained after full demineralisation (FD method). The nuclear DNA of the samples was quantified using the Investigator Quantiplex kit and STR typing was performed using the NGM kit to evaluate the performance of tested extraction methods. In the aged DNA samples, 64% of full profiles were obtained using the FD method. For the contemporary skeletal remains the performance of the ND method was closer to the FD method compared to the old skeletons, giving 58% of full profiles with the ND method and 71% of full profiles using the FD method. The extraction of DNA from only 0.05 g of bone or tooth powder using the AutoMate Express has proven highly successful in the recovery of DNA from old and contemporary skeletons, especially with the modified FD method. We believe that the results obtained will contribute to the possibilities of using automated devices for extracting DNA from skeletal remains, which would shorten the procedures for obtaining high-quality DNA from skeletons in forensic laboratories.</description><subject>Bone and Bones - chemistry</subject><subject>Bones</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA - isolation & purification</subject><subject>DNA extraction</subject><subject>DNA Fingerprinting</subject><subject>Efficiency</subject><subject>Forensic Genetics - methods</subject><subject>Forensic sciences</subject><subject>History, 20th Century</subject><subject>Human remains</subject><subject>Humans</subject><subject>Microsatellite Repeats</subject><subject>Missing person identification</subject><subject>Pathology</subject><subject>Real-Time Polymerase Chain Reaction</subject><subject>Second World War</subject><subject>Skeletal remains</subject><subject>Skeletal system</subject><subject>Teeth</subject><subject>Tooth - chemistry</subject><issn>1752-928X</issn><issn>1878-7487</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kU1rFTEUhgdRbK3-ARcScONmpvmaZAZEKPWjQqkLFd2FTHJGM81MrkmmeP-9GW5V6KKrnMXzvuQ8p6qeE9wQTMTp1EyjnxuKSdsQ0mBMHlTHpJNdLXknH5ZZtrTuaff9qHqS0oRxy6lsH1dHVAjScsmPq28X7sdPv0cwjs44WDLSaw6zzmAR_M5Rm-zCgsKI3l6doTGGGQVvkV4sMmHJMO9C1HGP0jV4yNqjCLN2S3paPRq1T_Ds9j2pvr5_9-X8or789OHj-dllbTjrcz1YzKy0Wtu-h44xO7ABt1IYwg0XA_Sj0JwTotthkMxwOlIs8NAJpgeChWYn1atD7y6GXyukrGaXDHivFwhrUkUB6XvGel7Ql3fQKaxxKb8rlMAdL0JkoeiBMjGkFGFUu-jmsqIiWG3a1aQ27WrTrghRRXsJvbitXocZ7L_IX88FeH0AoLi4cRBV2mwbsC6CycoGd3__mztx493ijPbXsIf0fw-VqMLq83b47e6kxZhiydgf5oOoYQ</recordid><startdate>20160101</startdate><enddate>20160101</enddate><creator>Zupanič Pajnič, Irena</creator><creator>Debska, Magdalena</creator><creator>Gornjak Pogorelc, Barbara</creator><creator>Vodopivec Mohorčič, Katja</creator><creator>Balažic, Jože</creator><creator>Zupanc, Tomaž</creator><creator>Štefanič, Borut</creator><creator>Geršak, Ksenija</creator><general>Elsevier Ltd</general><general>Churchill Livingstone Inc., Medical Publishers</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20160101</creationdate><title>Highly efficient automated extraction of DNA from old and contemporary skeletal remains</title><author>Zupanič Pajnič, Irena ; Debska, Magdalena ; Gornjak Pogorelc, Barbara ; Vodopivec Mohorčič, Katja ; Balažic, Jože ; Zupanc, Tomaž ; Štefanič, Borut ; Geršak, Ksenija</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c439t-bd03d7daad99e833db3b0576c14c46be9f6a4411a5bb73c42f2060b863ab106a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Bone and Bones - chemistry</topic><topic>Bones</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA - isolation & purification</topic><topic>DNA extraction</topic><topic>DNA Fingerprinting</topic><topic>Efficiency</topic><topic>Forensic Genetics - methods</topic><topic>Forensic sciences</topic><topic>History, 20th Century</topic><topic>Human remains</topic><topic>Humans</topic><topic>Microsatellite Repeats</topic><topic>Missing person identification</topic><topic>Pathology</topic><topic>Real-Time Polymerase Chain Reaction</topic><topic>Second World War</topic><topic>Skeletal remains</topic><topic>Skeletal system</topic><topic>Teeth</topic><topic>Tooth - chemistry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zupanič Pajnič, Irena</creatorcontrib><creatorcontrib>Debska, Magdalena</creatorcontrib><creatorcontrib>Gornjak Pogorelc, Barbara</creatorcontrib><creatorcontrib>Vodopivec Mohorčič, Katja</creatorcontrib><creatorcontrib>Balažic, Jože</creatorcontrib><creatorcontrib>Zupanc, Tomaž</creatorcontrib><creatorcontrib>Štefanič, Borut</creatorcontrib><creatorcontrib>Geršak, Ksenija</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of forensic and legal medicine</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zupanič Pajnič, Irena</au><au>Debska, Magdalena</au><au>Gornjak Pogorelc, Barbara</au><au>Vodopivec Mohorčič, Katja</au><au>Balažic, Jože</au><au>Zupanc, Tomaž</au><au>Štefanič, Borut</au><au>Geršak, Ksenija</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Highly efficient automated extraction of DNA from old and contemporary skeletal remains</atitle><jtitle>Journal of forensic and legal medicine</jtitle><addtitle>J Forensic Leg Med</addtitle><date>2016-01-01</date><risdate>2016</risdate><volume>37</volume><spage>78</spage><epage>86</epage><pages>78-86</pages><issn>1752-928X</issn><eissn>1878-7487</eissn><abstract>Abstract We optimised the automated extraction of DNA from old and contemporary skeletal remains using the AutoMate Express system and the PrepFiler BTA kit. 24 Contemporary and 25 old skeletal remains from WWII were analysed. For each skeleton, extraction using only 0.05 g of powder was performed according to the manufacturer's recommendations (no demineralisation – ND method). Since only 32% of full profiles were obtained from aged and 58% from contemporary casework skeletons, the extraction protocol was modified to acquire higher quality DNA and genomic DNA was obtained after full demineralisation (FD method). The nuclear DNA of the samples was quantified using the Investigator Quantiplex kit and STR typing was performed using the NGM kit to evaluate the performance of tested extraction methods. In the aged DNA samples, 64% of full profiles were obtained using the FD method. For the contemporary skeletal remains the performance of the ND method was closer to the FD method compared to the old skeletons, giving 58% of full profiles with the ND method and 71% of full profiles using the FD method. The extraction of DNA from only 0.05 g of bone or tooth powder using the AutoMate Express has proven highly successful in the recovery of DNA from old and contemporary skeletons, especially with the modified FD method. We believe that the results obtained will contribute to the possibilities of using automated devices for extracting DNA from skeletal remains, which would shorten the procedures for obtaining high-quality DNA from skeletons in forensic laboratories.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>26615474</pmid><doi>10.1016/j.jflm.2015.11.001</doi><tpages>9</tpages></addata></record> |
fulltext | fulltext |
identifier | ISSN: 1752-928X |
ispartof | Journal of forensic and legal medicine, 2016-01, Vol.37, p.78-86 |
issn | 1752-928X 1878-7487 |
language | eng |
recordid | cdi_proquest_miscellaneous_1751993394 |
source | MEDLINE; Access via ScienceDirect (Elsevier) |
subjects | Bone and Bones - chemistry Bones Deoxyribonucleic acid DNA DNA - isolation & purification DNA extraction DNA Fingerprinting Efficiency Forensic Genetics - methods Forensic sciences History, 20th Century Human remains Humans Microsatellite Repeats Missing person identification Pathology Real-Time Polymerase Chain Reaction Second World War Skeletal remains Skeletal system Teeth Tooth - chemistry |
title | Highly efficient automated extraction of DNA from old and contemporary skeletal remains |
url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-16T23%3A34%3A55IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Highly%20efficient%20automated%20extraction%20of%20DNA%20from%20old%20and%20contemporary%20skeletal%20remains&rft.jtitle=Journal%20of%20forensic%20and%20legal%20medicine&rft.au=Zupani%C4%8D%20Pajni%C4%8D,%20Irena&rft.date=2016-01-01&rft.volume=37&rft.spage=78&rft.epage=86&rft.pages=78-86&rft.issn=1752-928X&rft.eissn=1878-7487&rft_id=info:doi/10.1016/j.jflm.2015.11.001&rft_dat=%3Cproquest_cross%3E3935299351%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1760844747&rft_id=info:pmid/26615474&rft_els_id=S1752928X15002073&rfr_iscdi=true |