Design and fabrication of an on-chip PCR device

Maybery, G. H. C. (2019-04)

Thesis (MEng)--Stellenbosch University, 2019.

Thesis

ENGLISH ABSTRACT: An investigation into microfluidic-based PCR (Polymerase Chain Reaction devices was conducted. A continous-flow serpentine-channel device was fabricated. Channels were etched on a Poly(methyl methacrylate (PMMA substrate using CO2 laser ablation and channels with dimensions 150 x 150µm were obtained. A PMMA lid was bonded via thermal-assisted solvent bonding using Isopropanol. An in-house external connection schema was fabricated for fluid inlet and outlet ports and pressure driven flow was achieved using a benchtop syringe pump. Notable amplification of a 250bp ( NisinA fragment was achieved within 30 minutes, a final concentration of 1.5 µg/L was detected. The relationship between flow rates, temperature zone residency times and amplification efficiency was examined. This device serves as a proof-ofconcept for two purposes, to show that fabrication of microfluidic devices can be accomplished in low-tech lab spaces and to provide a roadmap towards the design of a fully-portable, low cost PCR device.

AFRIKAANSE OPSOMMING: 'n Ondersoek na mikro-uidiese-gebaseerde PCR (Polimerase Kettingreaksie toestelle was uitgevoer. 'n Deurlopende-serpentine-kanaal toestel was vervaardig. Kanale was geëts op 'n Poly (methylmethacrylaat (PMMA substraat met CO2 laser ablasie en kanale met afmetings 150x150µm was behaal. 'n PMMA-deksel was gebind deur middel van hitte-geassisteerde oplosmiddel bindinggebruik Isopropanol. 'N interne eksterne konneksie skema was vervaardig vir inlaat en uitlaat poorte en druk gedrewe was behaal met behulp van a spuitbuisprop. Merkbare amplifikasie van 'n 250bp (NisinA fragment was binne 30 minute bereik, 'n nal konsentrasie van 1.5µg/L was opgespoor. Die verhouding tussen ow tariewe, temperatuursone verblyf tye en amptelike kategese was ondersoek. Hierdie toestel dien as 'n bewys van konsepte vir twee doeleindes, om aan te toon dat vervaardiging van mikro uidiese toestelle kan word behaal in lae-tegnologie laboratoriumruimtes en 'n padkaart na die ontwerp van 'n vol draagbare, laekoste-PCR-toestel.

Please refer to this item in SUNScholar by using the following persistent URL: http://hdl.handle.net/10019.1/105827
This item appears in the following collections: