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Microfluidic sample preparation forbacterial DNA assays
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Biology Education Centre. Ångströmlaboratoriet. (EMBLA)
2023 (English)Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

The aim of this project was to use microfluidics for diagnostics by taking a pre-existing sample preparation protocol and transferring it on-chip. By doing this the expectation was to reduce the total assay time in order to make the process of diagnostics quicker. Previous research has shown that transferring the protocol to on-chip would be possible since agarose allows for diffusion of the required chemicals and DNA fragments.

First part of the project was to design and build a microfluidic chip that could contain bacteria encapsulated in an agarose plug. The chip was built out of PDMS, glass and 3D printed parts containing the plug in the center of the chip. 

Second part of the project was to transfer the protocol on-chip and optimize it. Here, the bacteria inside the 55μL agarose plug were chemically lysed by connecting the chip to a peristaltic pump which allowed the chemicals to flow through the chip in a closed system. By changing parameters like total plug volume, temperature and flow rate the total assay time decreased from 4 hours to 1.5 hours. This gave a 62.5% reduction in total assay time from the original protocol.  

In conclusion, changing the original sample preparation protocol to a microfluidic approach significantly reduced the total assay time. Results also showed that it is possible for further optimization to be done which may lead to even more reduction in time. Another possible parameter which could be further optimized was the total plug volume which had potential to be further reduced. 

Place, publisher, year, edition, pages
2023. , p. 30
National Category
Engineering and Technology
Identifiers
URN: urn:nbn:se:uu:diva-504569OAI: oai:DiVA.org:uu-504569DiVA, id: diva2:1767546
Educational program
Master Programme in Applied Biotechnology
Supervisors
Examiners
Available from: 2023-08-07 Created: 2023-06-14 Last updated: 2023-08-07Bibliographically approved

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