Talk abstracts

Talk on Saturday 11:15-11:30am submitted by Collin Wetzel

Targeted Tandem Mass Spectrometry for tRNA Identification

Collin Wetzel (Department of Chemistry, University of Cincinnati), Patrick Limbach (Department of Chemistry, University of Cincinnati)

Abstract:
The global identification of individual transfer ribonucleic acids (tRNAs) can prove to be very difficult, requiring rigorous sample preparation prior to analysis. Here, we illustrate a highly optimized MS/MS assay for rapid global identification of tRNAs, building on techniques previously introduced by our group. The goal is to identify sequences of oligonucleotides that are unique to a certain isoacceptor tRNA by detecting unique fragmentation channels. This approach enables accurate tRNA identification in a manner more conducive to high-throughput analyses. Our results show 43 out of the 47 isoaccepting tRNAs predicted to be present in E. coli are routinely detected by targeting 22 precursor ions in less than 15 minutes. Validation experiments included the examination of tRNAs from a tRNA over-expression system. The approach is general enough that it could be applied to archaeal and eukaryotic organisms, and this approach should also be feasible for the targeted analysis of other small RNAs in the cell.

Keywords: tRNA, Reverse phase chromatography, Tandemn Mass Spectrometry