AN162: Determination of Nucleotides by Ion Chromatography with UV Absorbance Detection.

Instrument Type: IC

The purpose of this application note is to provide a simple and direct assay of nucleotides and deoxynucleotides in amplification cocktails used for PCR, rolling circle amplifications, DNA and RNA polymerase reactions, and reverse-transcriptase reactions. Nucleotides are separated by anion exchange on the DNAPac PA100 column using gradient elution and detected by absorbance at 260 nm. Resolution of these components provides a means to assay their quality and help identify causes of amplification failure.