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Multi- and Single-cellular Encapsulation within Microchannels for Effective Cell Lysis and DNA Extraction and Purification

Hassani Gangaraj, Mojtaba | 2021

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  1. Type of Document: M.Sc. Thesis
  2. Language: Farsi
  3. Document No: 53955 (08)
  4. University: Sharif University of Technology
  5. Department: Mechanical Engineering
  6. Advisor(s): Shamloo, Amir
  7. Abstract:
  8. In this study a droplet-based microfluidic system is desighned and fabricated to effectiverly lyse MCF7 cells and extract and purify their DNAs. The main purpose of this study is to transfer all the steps from macro scale to a microfluidic system containing a fluidic chip. This system is a semi automatic system and every part of the lysis and purification process is performed in one step. The first step is to encapsulate single cells and multi cells inside the droplets. By controlling the concentration of the cell solution, the number of encapsulated cells inside the droplet is efficiently and easily controlled and the cells were encapsulated as single cells and as multi cells inside the droplets. In the next step, the cells and the lysis reagent are simultanously encapsulated within droplets. Secondary flows inside the droplets were used to improve the mixing efficiency. The droplets containing the cell then enter the extraction and purification process. The droplets first go through two thermal processes, 65 and zero degrees and then enter the microchip designed for extraction. The purified DNAs are then extracted from the microchip and stored in a suitable environment. The final result is the DNA solution extracted from MCF-7 cells. The volume of the final solution is 100 microliter and based on the results obtained from nanodrop test, the concentration of the final solution is 4.3 Nano gram per microliter
  9. Keywords:
  10. Purification ; DNA Extraction ; Cell Lysis ; Single Cell Encapsulation ; Multicell Encapsulation ; Cancer Cells

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