Recent publications have outlined hybridization of a biotin probe to siRNA or ASO oligonucleotides followed by LC/MS analysis has produced lower LLOQ's than previously reported with either LLE or SPE extraction techniques. Here we took the same approach and applied it to PMO oligonucleotides. We also looked at probe design to better optimize probe design by taking into consideration the potential interference from the capture probe back to the oligonucleotide being quantitated in the assay along with accounting for optimal hybridization efficiency and melting off the oligonucleotide of interest from the streptavidin bead.
Learning Objectives:
Oligonucleotide hybridization and LC/MS analysis for PMO's and probe design.
Learn how to avoid interference from probes used in oligonucleotide hybridization to LC/MS analysis
Learn the basic KingFisher workflow for oligonucleotide hybridization
