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Study on Microbiochip for Buccal Cell Lysis and DNA Purification

상피세포 시료 전처리용 마이크로바이오칩에 관한 연구

  • Ha, Seung-Mo (Dept. of Mechanical Engineering, Hanyang Univ.) ;
  • Cho, Woong (Dept. of Mechanical Engineering, Hanyang Univ.) ;
  • Ahn, Yoo-Min (Dept. of Mechanical Engineering, Hanyang Univ.) ;
  • Hwang, Seung-Yong (Division of Molecular and Life Science, Hanyang Univ.)
  • 하승모 (한양대학교 기계공학과) ;
  • 조웅 (한양대학교 기계공학과) ;
  • 안유민 (한양대학교 기계공학과) ;
  • 황승용 (한양대학교 분자생명과학부)
  • Received : 2009.07.13
  • Accepted : 2010.10.10
  • Published : 2010.12.01

Abstract

This paper describes a separable microfluidic device fabricated with PDMS (polydimethylsiloxane) and glass. The device is used for sample preparation involving cell lysis and the DNA purification process. The cell lysis was performed for 2 min at $80^{\circ}C$ in a serpentine-type microreactor ($20 {\mu}l$) using a Au microheater that was integrated with a thermal microsensor on a glass substrate. The DNA that was mixed with other residual products during the cell lysis process was then filtered through a new filtration system composed of microbeads (diameter: $50 {\mu}m$) and PDMS pillars. Since the entire process (sample loading, cell lysis reaction, DNA purification, and sample extraction) was performed within 5 min in a microchip, we could reduce the sample preparation time in comparison with that for the conventional methods used in biochemistry laboratories. Finally, we verified the performance of the sample preparation chip by conducting PCR (polymerase chain reaction) analysis of the chip product.

중합효소 연쇄반응(PCR)을 수행하려면 세포 용해(cell lysis)와 DNA추출(DNA purification)과정이 포함된 시료 전처리 과정을 거쳐야 한다. 종래의 시료 전처리 과정은 계면활성제와 같은 세포용해 버퍼를 이용하거나 열 또는 전기적 방법으로 세포막 파열을 유도하여 세포벽을 깬 후에 잔여물 처리과정을 거쳐 DNA를 추출하게 된다. 본 연구에서는 마이크로 비드와 PDMS 기둥을 이용한 필터가 있는 시료 전처리용 바이오칩을 설계 및 제작하였다. 또한 제작된 바이오칩을 사용하여 $80^{\circ}C$에서 2분간 세포용해를 수행하고 DNA를 추출하였다. 칩에서 전처리과정을 거친 시료내의 DNA농도와 순도를 측정하고 DNA PCR과 겔 전기영동을 통해 시료 전처리용 바이오칩의 성능을 평가하였다.

Keywords

References

  1. Elizabeth, Y. H., Mather, L and Bell, J. L., 2002, "MEMS-Based Sample Preparation for Molecular Diagnosis," Anal Bioanal Chem, Vol. 371, pp. 49-65.
  2. Schilling, E. A., Kamholz, A. E. and Yager, P., 2002, "Cell Lysis and Protein Extraction in a Microfluidic Device with Detection by a Fluorogenic Enzyme Assay," Analytical Chemistry, Vol. 74, pp. 1798-1804. https://doi.org/10.1021/ac015640e
  3. Lee, D. W and Cho, Y.-H., 2008, "A Continuous Electrical Cell Lysis Chip Using a DC Bias Voltage for Cell Disruption and Electroosmotic Flow," Trans. of the KSME (A), Vol. 32, No. 10, pp. 831-835.
  4. Ke, C., Kelleher, A.-M., Berney, H., Sheehan, M. and Mathewson, A., 2007, "Single Step Cell Lysis/PCR Detection of Escherichia Coli in an Independently Controllable Silicon Microreactor," Sensors and Actuators B, Vol. 120, pp.538-544 https://doi.org/10.1016/j.snb.2006.03.019
  5. De Mello, A. J. and Bread, N., 2003, "Dealing with Real Samples: Sample Pre-Treatment in Microfluidic Systems," Lab Chip, Vol. 3, pp. 11N-19N. https://doi.org/10.1039/b301019h
  6. Witek, M. A., Liopis, S. D., Wheatley, A., McCarley, R. L. and Soper, S. A., 2006, "Purification and Preconcentration of Genomic DNA from Whole Cell Lysates Using Photoactivated polycarbonate (PPC) Microfluidic Chips," Nucleidic Acids Research, Vol. 34, No. 10, Article No. e74.
  7. Lee, C.-Y., Lee, G.-B., Lin, J.-L., Huang, F.-C. and Liao, C.-S., 2005, "Integrated Microfluidic Systems for Cell Lysis, Mixing/Pumping and DNA Amplification," J. Micromech. Microeng. Vol. 15, pp. 1215-1223. https://doi.org/10.1088/0960-1317/15/6/011
  8. Yang, J., Liu, Y., Rauch, C. B., Stevens, R. L., Lui, R. H., Lenigk, R. and Grodzinski, P., 2002, "High Sensitivity PCR Assay in Plastic Micro Reactors," Lab Chip, Vol. 2, pp.179-187 https://doi.org/10.1039/b208405h
  9. Shin, Y. S., Cho, K. C., Lim, S. H., Chung, S., Park, S.-J., Chung, C., Han, D.-C. and Chang, J. K., 2003, "PDMS-Based Micro PCR Chip with Parylene Coating," J. Micromech. Microeng., Vol. 13, pp.768-774 https://doi.org/10.1088/0960-1317/13/5/332
  10. Cho, C.-H., Cho, W., Ahn, Y. and Hwang, S.Y., 2006, "PDMS/Glass Serpentine Microchannel Chip for PCR with Bubble Suppression in Sample Injection," Trans. of the KSME (A), Vol. 30, No. 10, pp. 1261-1268.