Volume reduction solid phase extraction of DNA from dilute, large-volume biological samples

Reedy, Carmen R.; Bienvenue, Joan M.; Coletta, Lisa; Strachan, Briony C.; Bhatri, Naila; Greenspoon, Susan; Landers, James P.

doi:10.1016/j.fsigen.2009.09.005

« Previous Next »Forensic Science International: Genetics
Volume 4, Issue 3 , Pages 206-212, April 2010

Volume reduction solid phase extraction of DNA from dilute, large-volume biological samples

Carmen R. Reedy
- Department of Chemistry, University of Virginia, Charlottesville, VA 22904, United States
Joan M. Bienvenue
- Armed Forces DNA Identification Laboratory, Rockville, MD 20850, United States
Lisa Coletta
- Department of Chemistry, University of Virginia, Charlottesville, VA 22904, United States
Briony C. Strachan
- Department of Chemistry, University of Virginia, Charlottesville, VA 22904, United States
Naila Bhatri
- Armed Forces DNA Identification Laboratory, Rockville, MD 20850, United States
Susan Greenspoon
- Virginia Department of Forensic Science, Richmond, VA 23219, United States
James P. Landers
- Department of Chemistry, University of Virginia, Charlottesville, VA 22904, United States
- Department of Mechanical Engineering, University of Virginia, Charlottesville, VA 22904, United States
- Department of Pathology, University of Virginia Health Science Center, Charlottesville, VA 22908, United States
- Corresponding author at: Department of Chemistry, University of Virginia, McCormick Road, P.O. Box 400319, Charlottesville, VA 22904, United States. Tel.: +1 434 243 8658; fax: +1 434 243 8852.

Received 9 June 2009; received in revised form 2 September 2009; accepted 4 September 2009. published online 12 October 2009.

Abstract

Microdevices are often designed to process sample volumes on the order of tens of microliters and cannot typically accommodate larger volume samples without adversely affecting efficiency and greatly increasing analysis time. However, dilute, large-volume biological samples are frequently encountered, especially in forensic or clinical laboratories. A microdevice, capable of efficiently processing 0.5–1mL samples has been developed for solid phase extraction (SPE) of DNA. SPE was carried out on a microdevice utilizing magnetic silica particles and an optimized volumetric flow rate and elution buffer, resulting in a 50-fold decrease in volume and a 15-fold increase in DNA concentration. Device characterization studies showed DNA extraction efficiencies comparable with previously reported silica-based purification methods, with robust performance demonstrated by the successful amplification of a fragment from the gelsolin gene extracted from dilute whole blood. In addition, the microchip-based method for SPE of large volume, dilute samples was also used to demonstrate the first successful on-chip purification of mitochondrial DNA (mtDNA) from both dilute whole blood and a degraded blood stain.