Volume reduction solid phase extraction of DNA from dilute, large-volume biological samples.
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 0.5-1 mL 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.
Reedy
Carmen R
CR
Department of Chemistry, University of Virginia, Charlottesville, VA 22904, USA.
Bienvenue
Joan M
JM
Coletta
Lisa
L
Strachan
Briony C
BC
Bhatri
Naila
N
Greenspoon
Susan
S
Landers
James P
JP
D001665:Binding Sites; D001769:Blood; D001801:Blood Stains; D004247:DNA / Q000032:analysis*; D004272:DNA, Mitochondrial / Q000032:analysis; D005453:Fluorescence; D006801:Humans; D007202:Indicators and Reagents; D046210:Microfluidic Analytical Techniques; D016133:Polymerase Chain Reaction; D012822:Silicon Dioxide; D052616:Solid Phase Extraction
D004272:DNA, Mitochondrial; D007202:Indicators and Reagents; D012822:Silicon Dioxide; D004247:DNA