Bio-lab Technique used in DNA computing

Cell & Microbial Engineering Lab. 1

Bio-lab Technique used in DNA computing

Jung-Ah KimSeoul National Uninversity

2002. 6. 5.


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


Bio-lab technique(1)

Separation Gel electrophoresis Polyacylamide gel electrophoresis Capillary electrophoresis Bead separation


Gel electrophoresis

Molecular size fraction technique

Detect the specific DNA

Bead Separation

Solution detection or filtering step!


Gel ElectrophoresisGel Electrophoresis


Complementary

Magnetic Beads

Magnet

Bead Separation (1)Bead Separation (1)


Bead Separation (2)Bead Separation (2)

Biotin (Vitamin H)


Bio-lab technique(2)

Detection RFLP SSCP DGGE TGGE SNP-PCR


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 .


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


Single base-pair difference at a specific site in the DNA

PCR 하는 동안 증폭된 특정유전자를 형광 hybridization probe 를 이용해 검출

SNP-PCR


DNA computin on surfaces(Liu et al.,Nature 403 ,143-144 ,2000)


Overvies of the surface-based approach to DNA computation


Encoding Scheme

Problem:


Four cycles of SAT computation

Destroyed using the single strand specific E.coli exonuclease I


Fluorescence profile & intensity histogram


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


Scheme showing behind electrical detection of DNA


Capture DNA strand

Target DNA strand

Probe DNA strand


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


Resistance of the electrode arrays measured as a function of increasing silver enhancing time


장점 : high sensitive(1:100,000)

PCR 불필요

mismatch 된 DNA 를 줄일 수 있다 .

단점 : variable 수가 적을 때에 유용


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

Bio-lab Technique used in DNA computing

Documents

Transcript of Bio-lab Technique used in DNA computing