Design and Fabrication of Microfluidic System for Cell Lysis and DNA Purification

Jalilvand, Elahe | 2022

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  1. Type of Document: M.Sc. Thesis
  2. Language: Farsi
  3. Document No: 55088 (08)
  4. University: Sharif University of Technology
  5. Department: Mechanical Engineering
  6. Advisor(s): Shamloo, Amir; Hosseini, Vahid
  7. Abstract:
  8. Nowadays, intracellular studies have been widely developed in biological applications. Intracellular analysis requires direct experiments on cells such as cell separation, purification, lysis and DNA extraction. The process of cell lysis and the resulting DNA purification is a crucial step in diagnostic processes. Since, the basis of many genetic studies is the information existed in double-stranded DNA structures.In this project, a microfluidic system is presented which performs integrated chemical cell lysis and DNA extraction. The cell used in this study is L-929. In order to increase cell lysis, the serpentine micromixer in combination with internal and external barriers at the beginning of the microchannel was designed to maximize the mixing of cell and buffer solutions. In order to further contact of two fluids, a sandwich mechanism is used in such way that the cellular solution enters the channel from the middle and buffer solution enters crosswise from both sides. After that, the fluid is heated in a reservoir with specific dimensions. Then, to complete the lysis process, absolute ethanol enters and all products pass through a silicon filter placed at the end of the channel and the DNA is trapped in this filter. In the next step, washing buffers 1 and 2 and elution buffer enter the channel, respectively and pass through the filter. The final output solution which has volume of about 1000 microliters, contains DNA. For investigating DNA quantity, a nanodrop test is carried out. It shows that the DNA is purified with suitable concentration (76/054ng⁄μL) and high purity (1/893). The results of experiment show that the developed device lyses the entrance cells with high efficiency (96/6%) and shorter time compared to conventional methods. Also, the purified DNA could be used in downstream applications like PCR
  9. Keywords:
  10. Microfluidic System ; Cell Lysis ; Micromixer ; DNA Purification ; Cell Separation