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CHARACTERIZATION OF AN OLIGONUCLEOTIDE-BINDING FLUORESCENT LIGAND FOR APPLICATION IN AFFINITY PURIFICATION OF DSDNA

TitleCHARACTERIZATION OF AN OLIGONUCLEOTIDE-BINDING FLUORESCENT LIGAND FOR APPLICATION IN AFFINITY PURIFICATION OF DSDNA
Publication TypeJournal Article
Year of Publication1995
AuthorsHaynes, CA, Sherwood, CS, Turner, RFB
JournalBiotechnology and Bioengineering
Volume48
Pagination25-35
Date PublishedOct
Type of ArticleArticle
ISBN Number0006-3592
KeywordsANTHRACYCLINE, antibiotics, BIO-POLYMERS, DNA AMPLIFICATION, DNA PURIFICATION, DNA-BINDING FLUOROPHORE, DYES, FLUORESCENT LIGAND, POLYMERASE CHAIN-REACTION, SENSITIVITY
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. (C) 1995 John Wiley & Sons, Inc.

URL<Go to ISI>://A1995RU62700004