(A) Schematic representation of multiplex quantification of 5′-tRNA halves.
(B) We used 10 sets of synthetic RNA mixtures to confirm the specificity of the multiplex TaqMan RT-qPCR method. The 5′-tRNAHisGUG half (HEX), 5′-tRNAGlyGCC half (TAMRA), and spike-in RNA (FAM) are simultaneously quantified in Group 1, and the 5′-tRNALysCUU half (HEX) and spike-in RNA (FAM) are quantified in Group 2.
(C) The multiplex method was applied to the 10 sets of synthetic RNAs. The results showed specific detection of targeted RNAs by each TaqMan probe without cross-reaction. Asterisk indicates that the amplification signal was not detected.