A high volume extraction and purification method for recovering DNA from human bone

Pamela L Marshall; Monika Stoljarova; Sarah E Schmedes; Jonathan L King; Bruce Budowle

doi:10.1016/j.fsigen.2014.06.011

A high volume extraction and purification method for recovering DNA from human bone

Forensic Sci Int Genet. 2014 Sep:12:155-60. doi: 10.1016/j.fsigen.2014.06.011. Epub 2014 Jun 18.

Authors

Pamela L Marshall¹, Monika Stoljarova², Sarah E Schmedes³, Jonathan L King³, Bruce Budowle⁴

Affiliations

¹ Institute of Applied Genetics, Department of Molecular and Medical Genetics, University of North Texas Health Science Center, 3500 Camp Bowie Blvd., Fort Worth, TX 76107, USA. Electronic address: pamela.marshall@unthsc.edu.
² Institute of Applied Genetics, Department of Molecular and Medical Genetics, University of North Texas Health Science Center, 3500 Camp Bowie Blvd., Fort Worth, TX 76107, USA; Centre for Biology of Integrated Systems, Tallinn University of Technology, Akadeemia tee, Tallinn, Estonia.
³ Institute of Applied Genetics, Department of Molecular and Medical Genetics, University of North Texas Health Science Center, 3500 Camp Bowie Blvd., Fort Worth, TX 76107, USA.
⁴ Institute of Applied Genetics, Department of Molecular and Medical Genetics, University of North Texas Health Science Center, 3500 Camp Bowie Blvd., Fort Worth, TX 76107, USA; Center of Excellence in Genomic Medicine Research (CEGMR), King Abdulaziz University, Jeddah, Saudi Arabia.

PMID: 24997320
DOI: 10.1016/j.fsigen.2014.06.011

Abstract

DNA recovery, purity and overall extraction efficiency of a protocol employing a novel silica-based column, Hi-Flow(®) (Generon Ltd., Maidenhead, UK), were compared with that of a standard organic DNA extraction methodology. The quantities of DNA recovered by each method were compared by real-time PCR and quality of DNA by STR typing using the PowerPlex(®) ESI 17 Pro System (Promega Corporation, Madison, WI) on DNA from 10 human bone samples. Overall, the Hi-Flow method recovered comparable quantities of DNA ranging from 0.8ng±1 to 900ng±159 of DNA compared with the organic method ranging from 0.5ng±0.9 to 855ng±156 of DNA. Complete profiles (17/17 loci tested) were obtained for at least one of three replicates for 3/10 samples using the Hi-Flow method and from 2/10 samples with the organic method. All remaining bone samples yielded partial profiles for all replicates with both methods. Compared with a standard organic DNA isolation method, the results indicated that the Hi-Flow method provided equal or improved recovery and quality of DNA without the harmful effects of organic extraction. Moreover, larger extraction volumes (up to 20mL) can be employed with the Hi-Flow method which enabled more bone sample to be extracted at one time.

Keywords: Bone; DNA quantity; Hi-Flow(®) silica column; Organic extraction; PCR inhibition; STR typing.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bone and Bones / chemistry*
DNA / isolation & purification*
Electrophoresis, Capillary
Humans
Microsatellite Repeats

Substances

DNA