High Volume Extraction and Purification Method for Recovering DNA from Human Bone

• NCJ Number: 248154
• Journal: Forensic Science International: Genetics Volume: 12 Dated: September 2014 Pages: 155-160
• Author(s): Pamela L. Marshall; Monika Stoljarova; Sarah E. Schmedes; Jonathan L. King; Bruce Budowle
• Date Published: June 2014
• Length: 6 pages
• Grant Number(s): 2009-DN-BX-K188; 2012-IJ-CX-0016
• Agencies: NIJ-Sponsored
• Publication Type: Research (Applied/Empirical)
• Award: "Development of an Analytical Approach for a Robust Probability Model for LCN DNA Profile Interpretation"; Improved Tools and Interpretation Guidelines for Examining Limited Low Copy Number DNA Obtained from Degraded Single Source Samples: Bones, Teeth, and Hairs

Annotation

This study assessed the DNA recovery, purity, and overall extraction efficiency of a novel silica-based column, Hi-Flow protocol in identifications from human bone, comparing its performance with that of a standard organic DNA extraction methodology.

Abstract

Overall, the general Hi-Flow method (Generon Ltd.,Maidenhead, UK) recovered similar quantities of DNA as the organic method for the majority of the bones studied; however, in contrast with the organic method, the Hi-Flow protocol reduced the number of handling steps (26 total steps compared with 20 total steps, respectively); and there were two sample transfers with the Hi-Flow method compared with five for the organic extraction procedure. This reduced the risk of cross-contamination. In addition, Hi-Flow extraction required only 4 hours of hands-on time compared with the organic extraction's 8 hours of hands-on time. Further, the Hi-Flow device enabled processing of samples in a larger volume because of an increased buffer and bone powder capacity.. This increases DNA yield compared to the organic method. The Hi-Flow protocol also allowed extraction in a single tube and reduced time and manipulation needed to perform purification; the sample size, however, was limited. Additional studies could increase understanding of the overall performance of increased amounts of bone and larger volume extraction. Another feature of the Hi-Flow method is the elimination of the use of hazardous compounds such as phenol and chloroform. Materials and methods are described. 2 tables, 1 figure, and 21 references