tRNA-Seq of Mtb in vivo: MtbH37RV total RNA at 0h rep1
To construct the library, RNA samples were ligated to the adenylated 3p-v4 adaptors, and reverse transcription was performed with ProtoScript II RT enzyme (NEB) using barcode primers. Finally, PCR amplification was performed with tRNAs oligoFor mix and A-PE-PCR10 (after 5 cycles, the program was paused and B_i7RPI1_CGTGAT or i7RPI7_ GATCTG was added) using Q5 Polymerase Hot-Start (NEB). The library was sequenced by DNBSEQ -G400RS High-throughput Sequencing Set (PE100) in BGI Genomics (Hong Kong).