title: Pre-storage of liquid reagents in glass ampoules for DNA extraction on a fully integrated lab-on-a-chip cartridge.
abstract: Self-containing, ready-to-use cartridges are essential for mobile Lab-on-a-Chip (LoaC) systems intended for Point-of-Care (POC) use. Up to now, a common weak point in many LoaC developments is the need to dispense liquid reagents into the test cartridge before or during processing of the assay. To address this issue we have developed an efficient method for fusing liquid reagents into glass ampoules, which are then sealed into a centrifugally operated cartridge. For on-demand reagent release, the ampoules are disrupted through the flexible lid of the cartridge. Upon centrifugation, 98.7 microL out of 100 microL (CV = 2.5%) of the pre-stored contents are released into the microfluidic system. No liquid loss is observed for ethanol and H(2)O stored for 300 days at room temperature. Frozen storage is possible without damage to the ampoules. Applicability of this concept is demonstrated by performing a LoaC integrated DNA extraction after 140 days of reagent pre-storage. DNA yield from 32 microL of whole blood was up to 199 ng, which is 77% of an off-chip reference extraction. The presented approach allows the improvement of existing LoaC cartridges where pre-storage of liquid reagents was not implemented yet.
authors.affiliation: Laboratory for MEMS Applications, Department of Microsystems Engineering-IMTEK, University of Freiburg, Georges-Koehler-Allee 106, 79110 Freiburg, Germany.
authors.forename: Jochen
authors.initials: J
authors.lastname: Hoffmann
authors.forename: Daniel
authors.initials: D
authors.lastname: Mark
authors.forename: Sascha
authors.initials: S
authors.lastname: Lutz
authors.forename: Roland
authors.initials: R
authors.lastname: Zengerle
authors.forename: Felix
authors.initials: F
authors.lastname: von Stetten
mesh_terms: D004247:DNA / Q000737:chemistry* / Q000302:isolation & purification*; D005898:Glass; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D013048:Specimen Handling / Q000295:instrumentation*; D018511:Systems Integration
chemical_list: D004247:DNA