1. Integrated Microsystem of Isothermal Amplification of DNA and Electrophoresis on a Microfabricated Plastic Chip for Detection of Specific Gene and Analysis of Genetic Materials MEC Seminar Proteome Research Lab Proteome Research Lab 2003. 5. 20 Summarized by Park, Ji-Yoon Summarized by Park, Ji-Yoon Proceedings of the uTAS 2002 Symposium, Vol. 1, pp 215-217

2. Proteome Research Lab, http://gene.hanyang.ac.kr/ Proteome Research Lab, http://gene.hanyang.ac.kr/ 2002. 11. 27 Continuous-Flow PCR on Chip Science, 280, 1046-1048(1998) Fig 1. Schematic of a chip for flow through PCR 7

3. Proteome Research Lab, http://gene.hanyang.ac.kr/ Proteome Research Lab, http://gene.hanyang.ac.kr/ 2002. 11. 27 A Miniature Integrated Device for Automated Multiple Genetic Assays Nucleic Acids Research, 28, e60 (2000) Fig 4. An integrated device that automatically performs a multistep HIV genotyping assay 8

4. Proteome Research Lab, http://gene.hanyang.ac.kr/ Proteome Research Lab, http://gene.hanyang.ac.kr/ 2002. 11. 27 High Sensitivity PCR Assay in Plastic Micro Reactors Lab on a Chip, 2(4), 179-187(2002) 9

5. Proteome Research Lab, http://gene.hanyang.ac.kr/ Proteome Research Lab, http://gene.hanyang.ac.kr/ 2002. 11. 27 Infrared-mediated Thermocycling for DNA Amplification and Electrophoretic Separation on an Integrated Microrchip Device Proceedings of the uTAS 2002 Symposium, Vol. 1, pp 193-194 10 Fig 2. Microchip and thermocycling profile

6. Proteome Research Lab, http://gene.hanyang.ac.kr/ Proteome Research Lab, http://gene.hanyang.ac.kr/ 2002. 11. 27 DNA Amplification and Hybridization Assays in Integrated Plastic Monolithic Devices Proceedings of the uTAS 2002 Symposium, Vol. 1, pp 169-171 Fig 3. Monolithic integrated polycarbonated DNA assay device Anal. Chem. 74, 3063-3070 (2002) 11

7. Proteome Research Lab, http://gene.hanyang.ac.kr/ Proteome Research Lab, http://gene.hanyang.ac.kr/ 2002. 11. 27 On-chip integration of isothermal PCR and electrophoresis On-chip integration of isothermal PCR and electrophoresis Loop-mediated isothermal amplification(LAMP) Loop-mediated isothermal amplification(LAMP) Novel DNA amplification method developed by Eiken Chemical Co., Ltd. Novel DNA amplification method developed by Eiken Chemical Co., Ltd. By using 4 different primers specifically designed to recognize six distinct regions on the target gene By using 4 different primers specifically designed to recognize six distinct regions on the target gene Performed at a constant temperature using a strand displacement reaction. Performed at a constant temperature using a strand displacement reaction. Advantages Advantages High amplification efficiency( 10 9 -10 10 times in 15-60 minutes) High amplification efficiency( 10 9 -10 10 times in 15-60 minutes) High specificity High specificity The potential to replace PCR » simplicity, rapidity, specificity and cost-effectiveness The potential to replace PCR » simplicity, rapidity, specificity and cost-effectiveness Avoid the thermal cycling Avoid the thermal cycling Analysis Analysis by the integrated microchip electrophoresis by the integrated microchip electrophoresis by the naked eyes by adding fluorescent dye like SYBR Green I by the naked eyes by adding fluorescent dye like SYBR Green I Abstract 12

8. Proteome Research Lab, http://gene.hanyang.ac.kr/ Proteome Research Lab, http://gene.hanyang.ac.kr/ 2002. 11. 27 Loop-mediated Isothermal Amplification (LAMP) 13

9. Proteome Research Lab, http://gene.hanyang.ac.kr/ Proteome Research Lab, http://gene.hanyang.ac.kr/ 2002. 11. 27 RAM (Ramplification Amplification Method) 14

10. Proteome Research Lab, http://gene.hanyang.ac.kr/ Proteome Research Lab, http://gene.hanyang.ac.kr/ 2002. 11. 27 Chip Device and Electropherogram Integrated microchip for DNA amplification and CE Integrated microchip for DNA amplification and CE Reaction condition Reaction condition RAMP; 5- 10 ul volume, 50-65°C for 10-30min RAMP; 5- 10 ul volume, 50-65°C for 10-30min Micro-CE - Reproducibility(Fig. 4) Micro-CE - Reproducibility(Fig. 4) - Detection time; 10min (Fig 5) - Detection time; 10min (Fig 5) - Visualization(Fig 6) - Visualization(Fig 6) Fig 1. Structure of a PMMA chip and device and electropherogram 15

11. Proteome Research Lab, http://gene.hanyang.ac.kr/ Proteome Research Lab, http://gene.hanyang.ac.kr/ 2002. 11. 27 SYBR Green ™ I The dye (ex; EtBr) The dye (ex; EtBr) Binds preferentially to ds DNA & quantitate the amount of double- stranded product Binds preferentially to ds DNA & quantitate the amount of double- stranded product Stable to the extremes of temperature required for PCR reaction Stable to the extremes of temperature required for PCR reaction 6

12. Proteome Research Lab, http://gene.hanyang.ac.kr/ Proteome Research Lab, http://gene.hanyang.ac.kr/ 2002. 11. 27 The Reproducibility and Detection 16

13. Proteome Research Lab, http://gene.hanyang.ac.kr/ Proteome Research Lab, http://gene.hanyang.ac.kr/ 2002. 11. 27 Conclusions Integrated microsystem Integrated microsystem Useful for fast, sensitive, and simple detection of specific sequence of DNA Useful for fast, sensitive, and simple detection of specific sequence of DNA Application Application - O-157 E.coli, B. anthracis, Salmonella and other bacteria in the environment, food, animal, human blood, etc. - O-157 E.coli, B. anthracis, Salmonella and other bacteria in the environment, food, animal, human blood, etc. 17

14. Proteome Research Lab, http://gene.hanyang.ac.kr/ Proteome Research Lab, http://gene.hanyang.ac.kr/ 2002. 11. 27 Further Study Development of DNA Computing Chip Development of DNA Computing Chip DNA hybridization, ligation, PCR, CE DNA hybridization, ligation, PCR, CE the uTAS 2003 Symposium the uTAS 2003 Symposium Application of DNA Computing Application of DNA Computing Diagnosis, Gene expression, Environmental Checker etc… Diagnosis, Gene expression, Environmental Checker etc… uTAS 2002 Paper uTAS 2002 Paper To access the latest papers published in LOC please see To access the latest papers published in LOC please see http://www.rsc.org/is/journals/current/loc/lcadvarts.htm http://www.rsc.org/is/journals/current/loc/lcadvarts.htm http://www.rsc.org/is/journals/current/loc/lcadvarts.htm http://www.rsc.org/is/journals/current/loc/lcadvarts.htm For Advance articles http://www.rsc.org/is/journals/current/loc/lc002003.htm For Advance articles http://www.rsc.org/is/journals/current/loc/lc002003.htmhttp://www.rsc.org/is/journals/current/loc/lc002003.htm The general website is www.rsc.org/loc The general website is www.rsc.org/locwww.rsc.org/loc 18