Microwave-assisted digestion combined with silica-based spin column for DNA isolation from human bones

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dc.authoridOzdemir Kaynak, Elif/0000-0001-6667-8007
dc.authoridYesil-Celiktas, Ozlem/0000-0003-4509-2212
dc.contributor.authorOzdemir-Kaynak, Elif
dc.contributor.authorYesil-Celiktas, Ozlem
dc.date.accessioned2025-01-27T20:54:20Z
dc.date.available2025-01-27T20:54:20Z
dc.date.issued2015
dc.departmentÇanakkale Onsekiz Mart Üniversitesi
dc.description.abstractA protocol for the extraction of DNA from ancient skeletal material was developed. Bone specimen samples (powder or slice), buffer, pretreatment, and extraction methodologies were compared to investigate the best conditions yielding the highest concentration of DNA. The degree of extract contamination by polymerase chain reaction (PCR) inhibitors was compared as well. Pretreatment was carried out using agitation in an incubator shaker and microwave digestion. Subsequently, DNA from bones was isolated by the classical organic phenol-chloroform extraction and silica-based spin columns. Decalcification buffer for total demineralization was required as well as lysis buffer for cell lysis to obtain DNA, whereas microwave-assisted digestion proved to be very rapid, with an incubation time of 2 min instead of 24 h at an incubator shaker without using lysis buffer. The correction of isolated DNA was detected using real-time PCR with melt curve analysis, which was 82.8 +/- 0.2 degrees C for highly repetitive cc-satellite gene region specific for human chromosome 17 (locus D17Z1). Consequently, microwave-based DNA digestion followed by silica column yielded a high-purity DNA with a concentration of 19.40 ng/mu l and proved to be a superior alternative to the phenol-chloroform method, presenting an environmentally friendly and efficient technique for DNA extraction. (C) 2015 Elsevier Inc. All rights reserved.
dc.identifier.doi10.1016/j.ab.2015.06.034
dc.identifier.endpage50
dc.identifier.issn0003-2697
dc.identifier.issn1096-0309
dc.identifier.pmid26142220
dc.identifier.scopus2-s2.0-84937953228
dc.identifier.scopusqualityQ2
dc.identifier.startpage44
dc.identifier.urihttps://doi.org/10.1016/j.ab.2015.06.034
dc.identifier.urihttps://hdl.handle.net/20.500.12428/26046
dc.identifier.volume486
dc.identifier.wosWOS:000359881000009
dc.identifier.wosqualityQ2
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.indekslendigikaynakPubMed
dc.language.isoen
dc.publisherAcademic Press Inc Elsevier Science
dc.relation.ispartofAnalytical Biochemistry
dc.relation.publicationcategoryinfo:eu-repo/semantics/openAccess
dc.rightsinfo:eu-repo/semantics/closedAccess
dc.snmzKA_WoS_20250125
dc.subjectBones
dc.subjectDNA
dc.subjectInhibition
dc.subjectMicrowave-assisted digestion
dc.subjectPolymerase chain reaction
dc.titleMicrowave-assisted digestion combined with silica-based spin column for DNA isolation from human bones
dc.typeArticle

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