Microfluidic PCR devices with electrochemical detection of DNA

Atkins, Nigel Philip (2005) Microfluidic PCR devices with electrochemical detection of DNA. PhD thesis, University of Glasgow.

Abstract

This research undertaken involved designing, fabricating and testing of a
microfiuidic peR micro device with real-time electrochemical detection. The
aim was to provide an analytical device which could lower the cost and the
time taken for running DNA amplification. The later addition of automated
sample handling and detection would thereby reduce the time taken and
consequently the overall cost.
The real-time electrochemical detection utilised an electrochemical assay for
the detection of DNA invented by Molecular Sensing Ltd. It used a single
strand ferrocenylated probe DNA molecule which could be detected with an
electrochemical cell. The integration of an electrochemical cell was a key
feature of this work, along with the immobilisation of the Taq polymerase at
its working temperature.
Taq polymerase enzyme and T7 polymerase enzyme were immobilised on to
microspheres. Taq polymerase was immobilised in four ways and T7 polymerase
was immobilised in only one method. After immobilisation the enzymes
were unable to amplify DNA within peR experiments.
Microfiuidic peR devices, which incorporate the above two features, were
designed and fabricated. 3 basic ideas of devices were investigated, fiowthrough,
straight line and cyclic triangle device. All the devices had fundamental
problems which inhibited there ability to successfully amplify DNA.
An electrochemical assay was used within a microfiuidic device with internal
electrochemical detection, which utilised a filter to bring about sequence
specific DNA detection. Using biotinylated complementary probe DNA attached
to streptavidin coated beads to hybridise to the sample DNA. This
device incorporated a solid phase extraction and clean up step as well as
producing sequence specific DNA detection.

• Item Type: Thesis (PhD)
• Qualification Level: Doctoral
• Subjects: Q Science > QH Natural history > QH301 Biology; T Technology > TK Electrical engineering. Electronics Nuclear engineering
• Colleges/Schools: College of Science and Engineering > School of Engineering
• Supervisor's Name: Cooper, Professor Jon
• Date of Award: 2005
• Depositing User: Ms Mary Anne Meyering
• Unique ID: glathesis:2005-4880
• Copyright: Copyright of this thesis is held by the author.
• Date Deposited: 24 Jan 2014 09:56
• Last Modified: 24 Jan 2014 16:46
• URI: https://theses.gla.ac.uk/id/eprint/4880