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Updated: Jul 3, 2026

Using Modified Synthetic Oligonucleotides to Assay Nucleic Acid-Metabolizing Enzymes
Published on: July 5, 2024
Characterization of an oligonucleotide-binding fluorescent ligand for application in affinity purification of dsDNA
C A Haynes1, C S Sherwood, R F Turner
1Biotechnology Laboratory, The University of British Columbia, British Columbia, Canada.
Abstract:
The fluorescent probe PO-PRO-3 was investigated as a potential ligand for the affinity immobilization and purification of genomic or plasmid DNA fragments. Affinities and mechanisms for PO-PRO-3 binding to superhelical and linearized pUC 18 plasmid DNA were examined through measurement of binding isotherms, continuous-variation analysis, and DNA titrations. In addition, the effects of DNA conformation, protein and RNA contaminants, solvent polarity, and ionic strength are examined with the aim of optimizing binding and elution conditions and of assigning limits to the range of applicability of the affinity purification.

