title: Denaturing reversed-phase HPLC using a mobile phase containing urea for oligodeoxynucleotide analysis.
abstract: Denaturing reversed-phase (RP) high performance liquid chromatography (HPLC) is usually achieved by elevating column temperature. In this article, an alternative method involving using a mobile phase that contains urea and performing HPLC at room temperature is described. The efficacy of the new method was demonstrated by analyzing a 61-mer oligodeoxynucleotide (ODN) and double-stranded (ds) ODNs. The multiple peaks of the 61-mer ODN under normal conditions merged into one under the denaturing conditions. The broad single peaks of dsODNs under normal conditions were split into two sharp peaks.
authors: a Department of Chemistry , Michigan Technological University , Houghton , Michigan , USA.
authors: Suntara
authors: S
authors: Fueangfung
authors: Yinan
authors: Y
authors: Yuan
authors: Shiyue
authors: S
authors: Fang
mesh_terms: D001483:Base Sequence; D002021:Buffers; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D008969:Molecular Sequence Data; D009691:Nucleic Acid Denaturation / Q000187:drug effects; D009838:Oligodeoxyribonucleotides / Q000032:analysis* / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D013696:Temperature; D014508:Urea / Q000494:pharmacology*
chemical_list: D002021:Buffers; D009838:Oligodeoxyribonucleotides; D014508:Urea
keywords: DNA; Denaturing HPLC; oligodeoxynucleotide; urea