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Sequence‐Specific Oligonucleotide Purification Using Peptide Nucleic Acid Amphiphiles in Hydrophobic Interaction Chromatography
Author(s) -
Vernille James P.,
Schneider James W.
Publication year - 2004
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp049971z
Subject(s) - oligonucleotide , nucleic acid , amphiphile , peptide nucleic acid , chemistry , peptide , dna , hydrophilic interaction chromatography , chromatography , sequence (biology) , covalent bond , biochemistry , combinatorial chemistry , high performance liquid chromatography , organic chemistry , copolymer , polymer
We present a methodology to perform sequence‐specific separations of oligonucleotides using peptide nucleic acids covalently linked to alkane chains, or “PNA amphiphiles (PNAAs)”. The PNAA/DNA duplexes are discriminated from unbound DNA using hydrophobic interaction chromatography on a phenyl‐substituted Sepharose column. Nearly quantitative recovery is achieved at concentrations of 50 μM after incubation of oligomers with a stoichiometric amount of PNAA for 1 min or so. The method exhibits high sequence specificity, selectivity, and resolution when applied to mixtures of various oligomers up to 60 base pairs in length.