Separation of Nucleic Acids by Ion Pair Reversed Phase High Performance Liquid Chromatography (IP RP HPLC)

Ion-pair reversed-phase high performance liquid chromatography (IP RP HPLC) is shown to be an effective method to reliably analyze DNA. IP RP HPLC is a much faster and safer alternative to conventional methods of DNA separation and quantification. The method described here utilized a two-buffer effluent system consisting of triethylammonium acetate (TEAA) and acetonitrile (ACN). The method reliably separated and quantified DNA samples of 54 and 58 nt. This method will be used and optimized to separate similarly sized RNA samples. The ultimate goal is to separate mixtures of nucleotides generated from in-vitro transcription reactions