We demonstrate a microfluidic reaction chamber that mimics a microcentrifuge tube where reagents can be mixed sequentially at a known stoichiometry. The device has no moving parts or valves and is made by hot embossing in a polymer foil. Sample and reagents are filled in the reaction chamber by controlled guiding of the air/liquid interface in a rectangular array of pillars. The operation of the device is demonstrated by performing isothermal DNA amplification in nL volumes. In our device, 28 pg of DNA from λ-phage, a virus with a 48 kilo base genome, is amplified 500 times thus the amplification product is suitable for library preparation for second generation sequencing. We show that fabrication by hot embossing does not introduce significant contamination and that our device is performing comparably well to test tube amplification and current PDMS-based chip technology.
- DNA amplification