Sequence-specific oligonucleotide purification using peptide nucleic acid amphiphiles in hydrophobic interaction chromatography.
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 microM 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.
Vernille
James P
JP
Department of Chemical Engineering, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213-3890, USA.
Schneider
James W
JW
D001483:Base Sequence; D002847:Chromatography, Agarose / Q000379:methods*; D015342:DNA Probes / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D017403:In Situ Hybridization / Q000379:methods*; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry* / Q000302:isolation & purification*; D020135:Peptide Nucleic Acids / Q000737:chemistry*
D015342:DNA Probes; D009841:Oligonucleotides; D020135:Peptide Nucleic Acids