Table 1.

Non-specific reads in tRNA-seq approaches.

RNA reads	% non-tRNA (average)	species
YAMAT	3.2	Hsa
Pang-like procedure	43.5	Ddi, Gst
LOTTE-seq	3.0	Hsa, Sce, Ddi, Gst, Eco, Sol
RNA reads	% non-CCA (average)	species
YAMAT	6.3	Hsa
Pang-like procedure	58	Ddi, Gst
LOTTE-seq	0.6	Hsa, Sce, Ddi, Gst, Eco, Sol

The average number of sequence reads that did not map to tRNA genes (upper part) or that did not carry a 3ʹ-terminal CCA triplet (lower part) are shown for LOTTE-seq in comparison to YAMAT and a procedure closely related to the one described by Pang et al. For both criteria, LOTTE-seq shows the lowest amount of non-specific sequences. While YAMAT and LOTTE-seq show similarly low values for non-tRNA reads, adapter ligation by T4 DNA ligase is obviously more selective for nick sealing in CCA-sequence hybrids than the truncated T4 RNA ligase used in YAMAT. Due to the lack of CCA-specific adapter ligation, the Pang-like approach shows the highest amount of non-tRNA and non-CCA end ligation, illustrating the importance of the efficient separation of the tRNA fraction from other transcripts in the preparation procedure. Ddi, D. discoideum; Eco, E. coli; Gst, G. stearothermophilus; Hsa, HEK293T cell line; Sce, S. cerevisiae; Sol, S. oleracea.