A disposable, self-contained PCR chip.
A disposable, self-contained polymerase chain reaction (PCR) chip with on-board stored, just-on-time releasable, paraffin-passivated, dry reagents is described. During both storage and sample preparation, the paraffin immobilizes and protects the stored reagents. Fluid flow through the reactor leaves the reagents undisturbed. Prior to the amplification step, the chamber is filled with target analyte suspended in water. Upon heating the PCR chamber to the DNA's denaturation temperature, the paraffin melts and moves out of the way, and the reagents are released and hydrated. To better understand the reagent release process, a scaled up model of the reactor was constructed and the paraffin migration was visualized. Experiments were carried out with a 30 microl reactor demonstrating detectable amplification (with agarose gel electrophoresis) of 10 fg ( approximately 200 copies) of lambda DNA template. The in-reactor storage and on-time release of the PCR reagents reduce the number of needed operations and significantly simplifies the flow control that would, otherwise, be needed in lab-on-chip devices.
Kim
Jitae
J
Department of Mechanical Engineering and Applied Mechanics, University of Pennsylvania, Philadelphia, PA 19104-6315, USA.
Byun
Doyoung
D
Mauk
Michael G
MG
Bau
Haim H
HH
D003890:Desiccation; D004867:Equipment Design; D006361:Heating; D007202:Indicators and Reagents / Q000737:chemistry; D056656:Lab-On-A-Chip Devices; D046208:Microchip Analytical Procedures / Q000379:methods; D010232:Paraffin / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods
D007202:Indicators and Reagents; D010232:Paraffin