The analysis of mock DNA sequencing reactions using matrix‐assisted laser desorption/ionization mass spectrometry

Abstract

Abstract A mock sequencing experiment is described in which matrix‐assisted laser desorption/ionization (MALDI) mass spectrometry is used to analyze mixtures of synthetic oligodeoxynucleotides chosen to match the first 24 DNA fragments generated in typical sequencing reactions using a standard M13mp19 template and primer. The experiments permitted the evaluation of important issues relating to resolution, sensitivity, multiple charging, and adduct formation. Mass resolutions (full width at half maximum) were between 100 and 200, and it was relatively straightforward to assign bases in the sequencing simulation using subpicomolar quantities of material. These results demonstrate the potential of MALDI mass spectrometry in DNA sequence analysis.