Incorporation of spiral continuous flow (CF) polymerase chain reaction (PCR) devices into a 96-well titer plate format will enable highly parallel analyses of many nucleic acids under steady-state temperature control for the realization of a high throughput CFPCR multi-reactor platform. Prior to realization of the full platform, verifying the feasibility of small footprint CFPCR devices, each device confined to a footprint of 8x8 mm 2 , throughout the manufacturing and amplification processes was essential. Small footprint, CFPCR devices in a CFPCR multi-reactor module were designed and double-side hot embossed on one side of a substrate and structures for thermal isolation on the other to improve thermal management. Successful amplification of 99 bp target DNA fragments from λ-DNA template was demonstrated in the small footprint, 20-and 25-turn CFPCR devices.
ABSRACTA 96-well solid-phase reversible immobilization (SPRI) reactor in a micro molded, photo-activated polycarbonate (PPC) chip was designed, fabricated, and used to purify genomic DNA (gDNA) in a highly parallelized fashion. A SPRI reactor was designed to fit in a 96-well microtiter plate format with compatibility to the existing samples/reagents loading technologies. A high quality, metallic, large area mold insert (LAMI) was fabricated using a SU-8 based UV-LIGA technique. Micro molding of polycarbonate (PC) was done using hot embossing, resulting in good replication fidelity over the large surface area. The 96-well PPC-SPRI reactor, assembled by thermal fusion bonding, showed 100% efficiency for simple and fast purification of gDNA from whole cell lysates of bacterial species. The 96-well PPC-SPRI reactor will open up new avenues for low cost, disposable DNA sample purification in high throughput applications.
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