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Other Detection System of Theorem Proving
2004. 2. 10
Park, Ji-Yoon
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February 10, 2004 February 10, 2004 APRL http://gene.hanyang.ac.krAPRL http://gene.hanyang.ac.kr 22
3 × 3 Matrix Chip (1)
1 × 3 Matrix
3 × 3 Matrix
Poly-L-Lysine
45 mer15 mer
15 mer
45 mer15 mer
15 mer
45 mer15 mer
15 mer
15 m15 merer
15 m15 merer
45 m45 merer
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Experimental Steps
1. Poly-L-Lysine Coating 1. Poly-L-Lysine Coating
3. Washing & Dry 3. Washing & Dry
4. Attachment of Target DNA 4. Attachment of Target DNA
6. Washing & Dry 6. Washing & Dry
7. Confocal LIF Detection 7. Confocal LIF Detection
5. S1 nuclease Digestion 5. S1 nuclease Digestion
2. Attachment of Probe DNA 2. Attachment of Probe DNA
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Amine-Coated Slide glass (Corning)
FAM(ST~Q)
Cy3(~P)
Cy5(R)
~S
~T
~RPQ
Target DNA
Probe DNA
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Poly-L-Lysine Patterned Slide glass
FAM Cy5
Overlay
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Problems & Discussion
PDMS Chip Channel Cross-talking Large sample volume require (~ 10 ul/channel)
We’ll use fresh PDMS Chip for every experiments Using Thin PDMS Chip
Low Intensity of Cy3 Cy 3 → Alexa fluoro dye (Molecular Probe)
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More Consideration
Hybridization(6 strands)
Ligation Surface Filtration
Nil Detection
S, T, Q, ¬ P, R
¬ S, ¬ T, Q, P,
¬ R
5’ ST5’ ST ¬¬ QQ ¬¬ PP RR
QPQP ¬¬ R 5’R 5’¬¬ SS ¬¬ TT
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Nanoparticle-based Detection (2)Nature 382, 607-609 (1996)
15 m15 merer
15 m15 merer
45 m45 merer
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Experimental Step
1. Au modification with thiol-capped oligonucleotides - Note: Oligonucleotide: non-complementary
3-thiol-TACCGTTG 3’-thiol-TTTGCTGA AuAuAuAu
2. Assembly (700 nm: aggregation)
- Addition of linking DNA duplex
5’-ATGGCAACTATACGCGCTAG
3’-ATATGCGCGATCTCAGCAAA 33. Immediately Color change
- with naked eyes
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Fig 1.
(A) The UV-vis spectrum of oligonucleotide-modified 13-nm particles before (red line) and after (blue line) DNA-induced assembly
(B) Solutions of 13-nm-diameter Au particles before (cuvette A) and after (cuvette B) DNA-induced assembly
(c) After extended periods of time a polymeric precipitate forms and settles to the bottom of the cuvette (cuvette C)
Expected Results (1)
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Fig 2. TEM images of DNA-linked nanoparticle assemblies at early stages in the growth process
Expected Results (2)