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 journalArticlepeer-review

20 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 languageEnglish
Pages (from-to)155-160
Number of pages6
JournalForensic Science International: Genetics
Volume12
DOIs
StatePublished - Sep 2014

Keywords

  • Bone
  • DNA quantity
  • Organic extraction
  • PCR inhibition
  • STR typing

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