1. Cell & Microbial Engineering Lab.1 Bio-lab Technique used in DNA computing Jung-Ah Kim Seoul National Uninversity 2002. 6. 5.

2. Cell & Microbial Engineering Lab.2 Contents Bio-lab techniques PCR Separation & Detection … DNA computing on a chip A new detection method Array – based Electrical Detection of DNA with Nanopartical probes

3. Cell & Microbial Engineering Lab.3 Bio-lab technique(1) Separation Gel electrophoresis Polyacylamide gel electrophoresis Capillary electrophoresis Bead separation

4. Cell & Microbial Engineering Lab.4 Gel electrophoresis Molecular size fraction technique Detect the specific DNA Bead Separation Solution detection or filtering step!

5. Cell & Microbial Engineering Lab.5 Gel Electrophoresis

6. Cell & Microbial Engineering Lab.6 Complementary Magnetic Beads Magnet Bead Separation (1)

7. Cell & Microbial Engineering Lab.7 Bead Separation (2) Biotin (Vitamin H)

8. Cell & Microbial Engineering Lab.8 Bio-lab technique(2) Detection RFLP SSCP DGGE TGGE SNP-PCR

9. Cell & Microbial Engineering Lab.9 RFLP(Restriction Fragment Length Polymorphysm) SSCP(Single Strand Conformation Polymorphysm) - 제한효소의 인지위의 존재여부의 차이에 의한 절단된 단편의 크기가 다름을 관찰하는 방법 - 염기서열의 차이에 따라 입체구조의 차이를 갖게 되어 전기영동 상에서 이동 속도의 차이를 보임.

10. Cell & Microbial Engineering Lab.10 DGGE(Denaturing Gradient Gel Electrophoresis) - detect differences in the melting behavior of small DNA fragments TGGE(Temperatuer Gradient Gel Electorphoresis) - gradient of increasing temperature down the gel

11. Cell & Microbial Engineering Lab.11 Single base-pair difference at a specific site in the DNA PCR 하는 동안 증폭된 특정유전자를 형광 hybridization probe 를 이용해 검출 SNP-PCR

12. Cell & Microbial Engineering Lab.12 DNA computin on surfaces (Liu et al.,Nature 403,143-144,2000)

13. Cell & Microbial Engineering Lab.13 Overvies of the surface-based approach to DNA computation

14. Cell & Microbial Engineering Lab.14 Encoding Scheme Problem :

15. Cell & Microbial Engineering Lab.15 Four cycles of SAT computation Destroyed using the single strand specific E.coli exonuclease I

16. Cell & Microbial Engineering Lab.16 Fluorescence profile & intensity histogram

17. Cell & Microbial Engineering Lab.17 A new technique for detection Array – based Electrical Detection of DNA with Nanopartical probes (So-Jung Park et al.,Science 295,1503-1506, 2002)

18. Cell & Microbial Engineering Lab.18 Scheme showing behind electrical detection of DNA

19. Cell & Microbial Engineering Lab.19 Capture DNA strand Target DNA strand Probe DNA strand

20. Cell & Microbial Engineering Lab.20 FE-SEM image of the nanoparticle probe on an glass surface (a) Before silver depositon Afer a treatment with the enhancer soultion (b) 3min (c) 6min (d) 9min

21. Cell & Microbial Engineering Lab.21 Resistance of the electrode arrays measured as a function of increasing silver enhancing time

22. Cell & Microbial Engineering Lab.22 장점 : high sensitive(1:100,000) PCR 불필요 mismatch 된 DNA 를 줄일 수 있다. 단점 : variable 수가 적을 때에 유용

23. Cell & Microbial Engineering Lab.23 Referance Liu et al.,Nature 403,143-144,2000 So-Jung Park et al.,Science 295,1503-1506, 2002 Elghanian et al, Science 277,1078-1081,1997 Li et al., Nucleic Acid Research 30,1558-1562,2002 Taton at al.,Science 289,1757-1759,2000