Highly effective DNA extraction method for nuclear short tandem repeat testing of skeletal remains from mass graves

Jon Davoren, Daniel Vanek, Rijad Konjhodzić, John Crews, Edwin Huffine, Thomas J. Parsons

Research output: Contribution to journalArticle

84 Citations (Scopus)

Abstract

Aim: To quantitatively compare a silica extraction method with a commonly used phenol/chloroform extraction method for DNA analysis of specimens exhumed from mass graves. Methods: DNA was extracted from twenty randomly chosen femur samples, using the International Commission on Missing Persons (ICMP) silica method, based on Qiagen Blood Maxi Kit, and compared with the DNA extracted by the standard phenol/chloroform-based method. The efficacy of extraction methods was compared by real time polymerase chain reaction (PCR) to measure DNA quantity and the presence of inhibitors and by amplification with the PowerPlex 16 (PP16) multiplex nuclear short tandem repeat (STR) kit. Results: DNA quantification results showed that the silica-based method extracted on average 1.94 ng of DNA per gram of bone (range 0.25-9.58 ng/g), compared with only 0.68 ng/g by the organic method extracted (range 0.0016-4.4880 ng/g). Inhibition tests showed that there were on average significantly lower levels of PCR inhibitors in DNA isolated by the organic method. When amplified with PP16, all samples extracted by silica-based method produced 16 full loci profiles, while only 75% of the DNA extracts obtained by organic technique amplified 16 loci profiles. Conclusions: The silica-based extraction method showed better results in nuclear STR typing from degraded bone samples than a commonly used phenol/chloroform method.

Original languageEnglish (US)
Pages (from-to)478-485
Number of pages8
JournalCroatian Medical Journal
Volume48
Issue number4
StatePublished - Aug 1 2007

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Microsatellite Repeats
DNA
Silicon Dioxide
Chloroform
Phenol
Body Remains
Bone and Bones
Femur
Real-Time Polymerase Chain Reaction
Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Medicine(all)

Cite this

Davoren, J., Vanek, D., Konjhodzić, R., Crews, J., Huffine, E., & Parsons, T. J. (2007). Highly effective DNA extraction method for nuclear short tandem repeat testing of skeletal remains from mass graves. Croatian Medical Journal, 48(4), 478-485.
Davoren, Jon ; Vanek, Daniel ; Konjhodzić, Rijad ; Crews, John ; Huffine, Edwin ; Parsons, Thomas J. / Highly effective DNA extraction method for nuclear short tandem repeat testing of skeletal remains from mass graves. In: Croatian Medical Journal. 2007 ; Vol. 48, No. 4. pp. 478-485.
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Davoren, J, Vanek, D, Konjhodzić, R, Crews, J, Huffine, E & Parsons, TJ 2007, 'Highly effective DNA extraction method for nuclear short tandem repeat testing of skeletal remains from mass graves', Croatian Medical Journal, vol. 48, no. 4, pp. 478-485.

Highly effective DNA extraction method for nuclear short tandem repeat testing of skeletal remains from mass graves. / Davoren, Jon; Vanek, Daniel; Konjhodzić, Rijad; Crews, John; Huffine, Edwin; Parsons, Thomas J.

In: Croatian Medical Journal, Vol. 48, No. 4, 01.08.2007, p. 478-485.

Research output: Contribution to journalArticle

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Davoren J, Vanek D, Konjhodzić R, Crews J, Huffine E, Parsons TJ. Highly effective DNA extraction method for nuclear short tandem repeat testing of skeletal remains from mass graves. Croatian Medical Journal. 2007 Aug 1;48(4):478-485.