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Methods for sequencing oligoribonucleotides and RNA by high performance liquid chromatography.
Authors:H Komiya  K Nishikawa  K Ogawa  S Takemura
Abstract:Conditions were established for the separation and quantitative determination of ribonucleosides, mono- and oligo-ribonucleotides by high-performance liquid chromatography (HPLC) on columns of AS-Pellionex SAX and AL-Pellionex WAX. By combining a high-speed UV spectrum monitor with an HPLC apparatus, products of RNase digestions of oligonucleotides and 5S ribosomal RNA (rRNA) were identified by measuring their UV spectra under continuous solvent flow, and also from their retention times on the columns (positions of elution). It took only 10 to 30 min for one chromatography run and required less than 0.01 A260 unit of sample per nucleotide material in each peak.
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