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Oligonucleotide- templated nanoparticle assembly

Oligonucleotide- templated nanoparticle assembly. Fiona McKenzie. Summary of previous work. Locked Nucleic Acid more stable duplex more selective for complementary target can target double stranded DNA: triplex formation Methods of detection: UV-visible spectroscopy

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Oligonucleotide- templated nanoparticle assembly

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  1. Oligonucleotide-templated nanoparticle assembly Fiona McKenzie

  2. Summary of previous work • Locked Nucleic Acid • more stable duplex • more selective for complementary target • can target double stranded DNA: triplex formation • Methods of detection: • UV-visible spectroscopy • SERRS: ideal for multiplexed analysis Small, 2007, 3, 1866–1868,Chem. Commun., 2008, 2367–2369

  3. 514.5 nm laser excitation Using SERRS to monitor DNA hybridisation Target DNA ROX ITC Addition of isothiocyanate dye after oligo functionalisation SH Aggregation of nanoparticles upon target DNA addition: allows electromagnetic coupling resulting in higher SERRS signals nanoparticle plasmons in resonance with laser excitation and dye absorbance ChemCommun. 2009, 5757–5759

  4. Using SERRS to monitor DNA hybridisation + target DNA Addition of target results in aggregation: increased SERRS output ChemCommun. 2009, 5757–5759

  5. Different Dye Labels Versatility of labelling method illustrated by use of different dye labels with different means of surface attachment- useful for multiplexing ChemCommun. 2009, 5757–5759

  6. Triplex SERRS 3’ 5’ 5’ 25 °C 60 °C 25 °C 60 °C 25 °C 60 °C 25 °C • Hybridisation of probes to yield triplex structure also gives rise to SERRS • Reversible assembly manuscript in preparation

  7. Towards a “SERRS ruler” 3’ 3’ 3’ 3’ anomaly in trend 5’ 5’ 62 °C 5’ 5’ 5’ 5’ 5’ 5’ 41 °C 65 °C 1 65 °C 5 10 • SERRS intensity dependence on inter-particle distance investigated • Greater the distance, lower intensity manuscript in preparation

  8. SERRS analysis Effect of melting temperature on SERRS intensity 39 °C 54 °C 51 °C NaCl used to alter melting temperature 47 °C 44 °C 39 °C Formamide used to alter melting temperature indicates thermodynamic stability of DNA nanoparticle assembly has an effect on SERRS intensity as well as interparticle distance

  9. Mixed Metal SERRS silver nanoparticle probes gold nanoparticle probes Gold nanoparticles often used in preference due to well-established surface chemistry and improved stability Silver nanoparticles give rise to much greater surface enhancement factors Nanoscale, 2009, in press

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