Nanoparticle-basedNanoparticle-based Theorem Theorem ProvingProving

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The Target Problem

R is true !!R is true !! nilnil

, add R as, add R as

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Experimental Section

Preparation of Au nanoparticles

Synthesis & Purification of (alkanethiol)-modified oligonucleotides

Preparation of 3'- or 5'-(Alkanethiol)oligonucleotide-modified Au nanoparticles

Preparation and Characterization of Linker DNA

Characterization of DNA-Linked Au Nanoparticle Assembly

Transmission Electron Microscopy (TEM)

Calculations of UV-vis Spectra

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Primer Sequence(5’→3’) Length ModificationGC Content

( % )

ST¬QAAG CAG TAG CGA CCA ATT GAC

GCA AAT TGA CGT ACG TAC GCT GAA

45 mer None 46.7

¬RPQCAT ACA ATG AAC GCA GTC AAC

GCA AGG CAG TTC AGC GTA CGT ACG

45 mer None 51.1

¬P CTG CCT TGC GTT GAC 15 mer 5’-phosphate 60.0

R TGC GTT CAT TGT ATG 15 mer 3’-thiol 40.0

¬S TGG TCG CTA CTT 15 mer 3’-thiol 53.3

¬T TCA ATT TGC GTC AAT 15 mer 5’-phosphate 33.3

The Oligomer Sequence

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UV-vis Spectrum of DNA Linked Au Nanoparticle

(A)(A) (B)(B) (C)(C)

DNA-linked nanoparticle DNA-linked nanoparticle Assembly Assembly

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Further Work

15 mer15 mer 15 mer15 mer 45 mer45 mer

Assembly Assembly

S1 Nuclease Digestion S1 Nuclease Digestion

Selectivity Test Selectivity Test

Surface-basedSurface-based Theorem Proving Theorem Proving

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Surfaced-based Theorem Proving

15 mer15 mer15 mer15 mer 45 mer45 mer

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¬RPQ

¬RPQ

¬RPQ

¬T

¬T

¬T

¬S

¬S

¬S

P.C

P.C

P.C

N.C

N.C

N.C

R

¬RPQ

¬RPQ

¬RPQ

¬T

¬T

¬T

¬S

¬S

¬S

P.C

P.C

P.C

N.C

N.C

N.C

¬P

ST ¬Q

(a)

(b)

The schematic diagram of theorem proving before (a) and after (b) hybridization. (a) The immobilized probe DNA sequences on aldehyde-coated slide glass. (b) Molecular recognition properties by the target molecules between the probe sequences.

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Experimental Section

Preparation of probe DNA

Preparation of Target DNA

Hybridization probe & Target DNA

Washing

Confocal microscope detectionConfocal microscope detection

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100 pmol

ST¬QST¬Q

50 pmolTemp. 50°C Constant

Temp. 60°C Constant

¬RPQ¬RPQ ¬T¬T ¬S ¬S P.CP.C N.C N.C

R

¬¬P

ST¬ST¬Q

ST¬QST¬Q

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100 pmol

¬¬P

50 pmolTemp. 50°C Constant

Temp. 60°C Constant

¬RPQ¬RPQ ¬T¬T ¬S ¬S P.CP.C N.C N.C

R

¬¬P

ST¬ST¬Q

¬¬P

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100 pmol

RR

50 pmolTemp. 50°C Constant

Temp. 60°C Constant

¬RPQ¬RPQ ¬T¬T ¬S ¬S P.CP.C N.C N.C

R

¬¬P

ST¬ST¬Q

RR

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Discussion

• Confocal microscope 457~633 nm Detection range dye & Non-overlapping emission wavelength Eqipment condition is break down now Microarray scanner (futher exp.)

• False positive signal ¬¬P and ¬¬S: non-complementary sequence Why (?)

GC ratio