| Summary: | 碩士 === 國立嘉義大學 === 生物機電工程學系研究所 === 97 === Integrating cell lysis and DNA extending process into a micrototal analytical system is one critical step for the analysis of nucleic acids. This study demonstrates a microfluidic system for cell lysis and DNA extending using electroporation.
The purpose of the present work first develops cell lysis technique in microfluidic channel using short electric field pulse. The study is combined photolithography to frabricate the PDMS microfluidic device with electromanipulation to lysis the U937 cell. The PDMS microfluidic device is consisted of two parts, first part is cell lysis area: Cells are immobilized on micro PDMS structures of an insulating barrier to generate field constriction and then enhance the yield of the electroporation process. After cell lysis, YO-PRO-1 will transfer to cell to dyne the cell DNA, and the other part is DNA extending area: using the peristaltic pump let DNA flow to the rectangular micro-structures. The histones of chromatin are degraded when the concentration of proteinase K is 98.5μg/ml. This study is successfully demonstrated an integrated microfluidic device capable of performing on-line cell lysis and DNA extraction and extending. The experimental result shows the use of the conditions applied electric 100Vpp, 1 kHz, and using osmotic pressure environment (75mM), let the DNA released to the solution, when the DNA fluorescence enlarged, then take advantage of proteinase K and drive the DNA flow by peristaltic pump, we can successfully let DNA flows behind the rectangle on the micro-structure.
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