Detection of transfer RNAs (tRNAs) by their signature digestion products

CHED 288

Cathy Cherenfant, ccherenfant@flsouthern.edu, Mahmud Hossain, and Patrick Limbach. Department of Chemistry, University of Cincinnati, PO Box 210172, Cincinnati, OH 45221-0172
The detection of transfer ribonucleic acids (tRNAs) was determined using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight (MALDI-TOF) mass spectrometry for its demonstration of accuracy and sensitive mass determination. Total transfer RNAs mixtures were isolated by an organic and solid-phase extraction from yeast Saccharaomyces cerevisiae strain YPH499 grown in YEPD media. Commercial grade yeast tRNA (Ambion Inc.) and YPH499 were digested with endonuclease RNase T1 and A. Signature products were then detected by mass spectrometry and compared to theoretical data.