What is the Nucleoside Digestion Mix?

Ivan Correa:

The Nucleoside Digestion Mix is an optimized mixture of enzymes that provides a convenient one-step method to generate single nucleosides for quantitative analysis of DNA or RNA modifications. It digests single, double-stranded, or DNA-RNA hybrids, and tolerates a wide range of base and ribose modifications.

In the first step, genomic or synthetic DNA or RNA is converted into individual nucleotides by the mix nuclease activity. In the second step, the nucleotides are dephosphorylated by the action of a non-specific phosphatase.

Here I would like to highlight an example where the Nucleoside Digestion Mix was used to monitor the metabolic incorporation of azido modified nucleosides into cellular RNA. Total RNA from cells treated with N⁶-ethylazido adenosine (in green), N⁶-propylazido adenosine (in magenta), and 2´-azido adenosine (in light brown) was digested to ribonucleosides using the Nucleoside Digestion Mix and analyzed by LC-MS.

The digested RNA from wild-type cells is shown in red. The insert shows an expansion of the overlaid chromatograms demonstrating the specific detection of the chemically-modified nucleosides.

To learn more, please visit the Nucleoside Digestion Mix product page on neb.com, and download our latest application note.