Study, Optimization and Construction of a Microfluidic Gene Amplification Device by Using Thin Film Layer Method

Eslami, Sara Sadat | 2021

8 Viewed
  1. Type of Document: M.Sc. Thesis
  2. Language: Farsi
  3. Document No: 54125 (06)
  4. University: Sharif University of Technology
  5. Department: Chemical and Petroleum Engineering
  6. Advisor(s): Vosoughi, Manouchehr; Alemzadeh, Iran; Shamloo, Amir
  7. Abstract:
  8. Polymerase Chain Reaction (PCR) is a process in which a special piece of a gene is amplified millions of times over a short period. This method has been of paramount importance in different fields of research and has been applied for different applications. PCR requires thermal cycling, or repeated temperature changes between two or three discrete temperatures to amplify specific nucleic acid target sequences. To achieve such thermal cycling, conventional bench-top thermal cyclers generally use a metal heating block powered by Peltier elements or benefit from forced convection heat transfer. Due to the fact that these methods are time consuming, it seems that design and fabrication of a fast PCR device can be definitely useful for genomic testing at the point-of-care (POC) level in which the test itself, obtaining the results, and their analysis must be done as soon as possible. In this piece of research, a microfluidic PCR system has been proposed which benefits from plasmonic photothermal heating for thermal cycling. In this research, an optical cavity PCR is proposed which utilizes two thin film layers of gold on the top and bottom of the PCR chamber. This system enhances the temperature uniformity across the PCR chamber. The results show that the heating and cooling rates for 50 μl volume of the sample are 3.3 °C/s and 2.4 °C/s, respectively. Considering the energy consumption of the proposed system, which uses only a 10 W LED for heating the sample, one can easily understand the efficiency of such a method for this purpose. The proposed protocol for amplifying the target DNA is suitable due to the results of the gel-electrophoresis test which bears witness to the fact that the system is able to amplify the sample perfectly. The ongoing research is focused on fabricating a PCR system on a PDMS-based microfluidic chip by using photolithography in order to increase the ratio of gold-coated surface area of the PCR chamber to the volume of the PCR sample
  9. Keywords:
  10. Polymerase Chain Reaction (PCR) ; Microfluidic System ; DNA Molecule ; Plasmonic Photothermal Heating ; Thermal Cycle ; Film Coating

 Digital Object List