Detection and quantification of glycosylated queuosine modified tRNAs by acid denaturing and APB gels

Analysis of Q and glycosylated Q-modified tRNAs using biotinylated tRNA probes. (A) Acid denaturing gels and northern blot of individual, Q or glycosylated Q-modified tRNA. 0Q: total RNA from cells without Q-modification; 100Q: total RNA from cells with full Q-modification; 0Q:100Q: equal amount mixture of 0Q and 100Q total RNA. G: unmodified tRNA, Q: Q-modified tRNA, mQ: mannosyl-Q, gQ: galactosyl-Q. (B) Acid denaturing gels and northern blot of tRNA^Asp and tRNA^Tyr with (NaIO₄) and without (NaCl) periodate treatment. 0: total RNA from 0Q cells; 1, 24: total RNA from cells 1 and 24 h after the addition of queuine to 0Q cells. Asterisk (*) indicates an unknown tRNA^Tyr species. Total RNA was from MCF7 cells.