Quantitative and qualitative analysis of a microfluidic DNA extraction system using a nanoporous AlO(x) membrane.
A nanoporous aluminium oxide membrane was integrated into a microfluidic system designed to extract hgDNA (human genomic DNA) from lysed whole blood. The effectiveness of this extraction system was determined by passing known concentrations of purified hgDNA through nanoporous membranes with varying pore sizes and measuring the amount of hgDNA deposited on the membrane while also varying salt concentration in the solution. DNA extraction efficiency increased as the salt concentration increased and nanopore size decreased. Based on these results, hgDNA was extracted from whole blood while varying salt concentration, nanopore size and elution buffer to find the conditions that yield the maximum concentration of hgDNA. The optimal conditions were found to be using a low-salt lysis solution, 100 nm pores, and a cationic elution buffer. Under these conditions the combination of flow and ionic disruption were sufficient to elute the hgDNA from the membrane. The extracted hgDNA sample was analysed and evaluated using PCR (polymerase chain reaction) to determine whether the eluted sample contained PCR inhibition factors. Eluted samples from the microfluidic system were amplified without any inhibition effects. PCR using extracted samples was demonstrated for several genes of interest. This microfluidic DNA extraction system based on embedded membranes will reduce the time, space and reagents needed for DNA analysis in microfluidic systems and will prove valuable for sample preparation in lab-on-a-chip applications.
Kim
Jungkyu
J
Department of Bioengineering, University of Utah, Salt Lake City, UT 84112, USA. jungkyu.kim@utah.edu
Gale
Bruce K
BK
D000537:Aluminum Oxide / Q000737:chemistry*; D004247:DNA / Q000097:blood / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D049329:Nanostructures / Q000737:chemistry*; D016062:Porosity; D044366:Transition Temperature
D004247:DNA; D000537:Aluminum Oxide