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Locked Nucleic Acid - Novel DNA Anolog

Locked Nucleic Acid - Novel DNA Anolog. MEC Meeting Hanyang University Proteome Research Lab 2002. 12. 13 Park, Ji-Yoon. Nucleic Acid Analogs. Comparison of LNA and PNA. Locked DNA Analog(LNA). Novel chemical DNA Anolog Bicyclic nucleic acid

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Locked Nucleic Acid - Novel DNA Anolog

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  1. Locked Nucleic Acid- Novel DNA Anolog MEC Meeting Hanyang University Proteome Research Lab 2002. 12. 13 Park, Ji-Yoon

  2. Nucleic Acid Analogs

  3. Comparison of LNA and PNA

  4. Locked DNA Analog(LNA) • Novel chemical DNA Anolog • Bicyclic nucleic acid • A ribonucleoside is linked between the 2’ oxygen and the 4’ carbon atoms with a methylene unit

  5. General Properties • Base pairs with DNA and RNA • Obey standard WC base pairing rule • Exceptionally high thermal stability • Improved discrimination • Compatible with most enzymes • Predictable melting behaviour • Synthesis by standard phosphoramidite chemistry using DNA-synthesizers • mixed with DNA, RNA and other nucleic acid analogs • synthesized with e.g. aminolinkers, biotin, Cy dyes etc • water soluble(gel electrophoresis, ethanol precipitation)

  6. Experimental Parameters or Application

  7. Melting Temperature( Tm )

  8. Nearest Neighbor Thermodynamics

  9. Fluorescence Microscope for generating Melting Curve • Fluorescence measurement in real-time PCR instrument • Decrease in fluorescence intensity from specific binding to ds DNA

  10. How LNA Substitution Increase ∆Tm

  11. Comparison of LNA and DNA Stability

  12. Effect of Single LNA Substitution

  13. www.LNA-Tm.com

  14. Sensitivity Test LNA Capture Probes(50mer) vs DNA Capture Probe(50mer) < Hybridization condition> A) 65°C, in 3x SSC B) Hybridization: 70°C, in 3x SSC

  15. Specificity Test LNA Capture Probes(40mer) vs DNA Capture Probe(40mer) • Hybridization condition: 65°C, in 3x SSC • - Perfect match: 1-7 - Mismatch: 2-8, 3-9, 4-10, 5-11, 6-12

  16. Microfluidic Chip in Action

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