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Cell & Microbial Engineering Lab. 1
Bio-lab Technique used in DNA computing
Jung-Ah KimSeoul National Uninversity
2002. 6. 5.
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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
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Bio-lab technique(1)
Separation Gel electrophoresis Polyacylamide gel electrophoresis Capillary electrophoresis Bead separation
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Gel electrophoresis
Molecular size fraction technique
Detect the specific DNA
Bead Separation
Solution detection or filtering step!
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Gel ElectrophoresisGel Electrophoresis
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Complementary
Magnetic Beads
Magnet
Bead Separation (1)Bead Separation (1)
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Bead Separation (2)Bead Separation (2)
Biotin (Vitamin H)
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Bio-lab technique(2)
Detection RFLP SSCP DGGE TGGE SNP-PCR
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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 .
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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
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Single base-pair difference at a specific site in the DNA
PCR 하는 동안 증폭된 특정유전자를 형광 hybridization probe 를 이용해 검출
SNP-PCR
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DNA computin on surfaces(Liu et al.,Nature 403 ,143-144 ,2000)
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Overvies of the surface-based approach to DNA computation
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Encoding Scheme
Problem:
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Four cycles of SAT computation
Destroyed using the single strand specific E.coli exonuclease I
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Fluorescence profile & intensity histogram
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A new technique for detection Array –based Electrical Detection of DNA with Nanopartical probes (So-Jung Park et al.,Science 295,1503-1506, 2002)
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Scheme showing behind electrical detection of DNA
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Capture DNA strand
Target DNA strand
Probe DNA strand
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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
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Resistance of the electrode arrays measured as a function of increasing silver enhancing time
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장점 : high sensitive(1:100,000)
PCR 불필요
mismatch 된 DNA 를 줄일 수 있다 .
단점 : variable 수가 적을 때에 유용
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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