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

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

Research output: Contribution to journal › Article › peer-review

23 Scopus citations

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 0.8 ng ± 1 to 900 ng ± 159 of DNA compared with the organic method ranging from 0.5 ng ± 0.9 to 855 ng ± 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 20 mL) can be employed with the Hi-Flow method which enabled more bone sample to be extracted at one time.

Original language	English
Pages (from-to)	155-160
Number of pages	6
Journal	Forensic Science International: Genetics
Volume	12
DOIs	https://doi.org/10.1016/j.fsigen.2014.06.011
State	Published - Sep 2014

Keywords

Bone
DNA quantity
Organic extraction
PCR inhibition
STR typing

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10.1016/j.fsigen.2014.06.011

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title = "A high volume extraction and purification method for recovering DNA from human bone",

abstract = "DNA recovery, purity and overall extraction efficiency of a protocol employing a novel silica-based column, Hi-Flow{\textregistered} (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 {\textregistered} 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 0.8 ng ± 1 to 900 ng ± 159 of DNA compared with the organic method ranging from 0.5 ng ± 0.9 to 855 ng ± 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 20 mL) can be employed with the Hi-Flow method which enabled more bone sample to be extracted at one time.",

keywords = "Bone, DNA quantity, Organic extraction, PCR inhibition, STR typing",

author = "Marshall, {Pamela L.} and Monika Stoljarova and Schmedes, {Sarah E.} and King, {Jonathan L.} and Bruce Budowle",

note = "Funding Information: This project was supported in part by award nos. 2009-DN-BX-K188 and 2012-IJ-CX-0016 , awarded by the National Institute of Justice, Office of Justice Programs, U.S. Department of Justice . The opinions, findings, and conclusions or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect those of the Department of Justice. ",

year = "2014",

month = sep,

doi = "10.1016/j.fsigen.2014.06.011",

language = "English",

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journal = "Forensic Science International: Genetics",

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T1 - A high volume extraction and purification method for recovering DNA from human bone

AU - Marshall, Pamela L.

AU - Stoljarova, Monika

AU - Schmedes, Sarah E.

AU - King, Jonathan L.

AU - Budowle, Bruce

N1 - Funding Information: This project was supported in part by award nos. 2009-DN-BX-K188 and 2012-IJ-CX-0016 , awarded by the National Institute of Justice, Office of Justice Programs, U.S. Department of Justice . The opinions, findings, and conclusions or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect those of the Department of Justice.

PY - 2014/9

Y1 - 2014/9

N2 - 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 0.8 ng ± 1 to 900 ng ± 159 of DNA compared with the organic method ranging from 0.5 ng ± 0.9 to 855 ng ± 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 20 mL) can be employed with the Hi-Flow method which enabled more bone sample to be extracted at one time.

AB - 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 0.8 ng ± 1 to 900 ng ± 159 of DNA compared with the organic method ranging from 0.5 ng ± 0.9 to 855 ng ± 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 20 mL) can be employed with the Hi-Flow method which enabled more bone sample to be extracted at one time.

KW - Bone

KW - DNA quantity

KW - Organic extraction

KW - PCR inhibition

KW - STR typing

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

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