Calorimetric and Spectroscopic Analysis of the Thermal Stability of Short Duplex DNA-Containing Sugar and Base-Modified Nucleotides.

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  • Additional Information
    • Source:
      Publisher: Academic Press Country of Publication: United States NLM ID: 0212271 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1557-7988 (Electronic) Linking ISSN: 00766879 NLM ISO Abbreviation: Methods Enzymol Subsets: MEDLINE
    • Publication Information:
      Original Publication: New York, Academic Press.
    • Subject Terms:
      DNA/*chemistry ; Nucleotides/*chemistry ; Oligonucleotides/*chemistry; Calorimetry, Differential Scanning ; Nucleic Acid Conformation ; Thermodynamics
    • Abstract:
      Base- and sugar-modified analogs of DNA and RNA are finding ever expanding use in medicine and biotechnology as tools to better tailor structured oligonucleotides by altering their thermal stability, nuclease resistance, base-pairing specificity, antisense activity, or cellular uptake. Proper deployment of these chemical modifications generally requires knowledge of how each affects base-pairing properties and thermal stabilities. Here, we describe in detail how differential scanning calorimetry and UV spectroscopy may be used to quantify the melting thermodynamics of short dsDNA containing chemically modified nucleosides in one or both strands. Insights are provided into why and how the presence of highly stable base pairs containing modified nucleosides can alter the nature of calorimetry or melting spectroscopy data, and how each experiment must therefore be conducted to ensure high-quality melting thermodynamics data are obtained. Strengths and weaknesses of the two methods when applied to chemically modified duplexes are also addressed.
      (© 2016 Elsevier Inc. All rights reserved.)
    • Contributed Indexing:
      Keywords: DNA; DNA melting thermodynamics; Differential scanning calorimetry; Locked nucleic acid; UV spectroscopy
    • Accession Number:
      0 (Nucleotides)
      0 (Oligonucleotides)
      9007-49-2 (DNA)
    • Publication Date:
      Date Created: 20160123 Date Completed: 20161017 Latest Revision: 20161230
    • Publication Date:
      20231215
    • Accession Number:
      10.1016/bs.mie.2015.08.001
    • Accession Number:
      26794352