Transcriptome, M.smegmatis MC2 155, 3 replicate, furan-2-carboxylate Cu with phenanthroline
Total RNA (1.0 - 2.5 g) was used to prepare the library. The quality of the obtained RNA was checked by microfluidic analyzer BioAnalyzer 2100 using the RNA 6000 pico chip kit (Agilent Technologies). Removal of residual genomic DNA from the samples was performed using the TURBO DNA-free Kit (Invitrogen). Ribosomal RNA (16S and 23S) was removed with the Ribo-Zero Plus rRNA Depletion Kit (Illumina). RNA purification was performed with the RNA Clean XP kit (Beckman Coulter). Genomic libraries were prepared with the NEBNext Ultra II Directional (NEB) kit. The size of the libraries was estimated by microfluidic analyzer BioAnalyzer 2100 using a High Sensitivity DNA chip (Agilent Technologies). The libraries were quantified by Qubit 4.0 Fluorometer (Thermo Scientific) using the Quant-iT DNA Assay Kit, High Sensitivity (Thermo Scientific). The resulting libraries were mixed in equimolar amounts. The quality of the finished libraries was assessed using a microfluidic analyzer BioAnalyzer 2100 (Agilent) using the Agilent DNA High Sensitivity Kit. Sequencing of the obtained libraries (diluted to 12 pM) was performed on a HiSeq 2500, using HiSeq Rapid SBS Kit v2 (200) (Illumina) (for paired reads 2  100 bp with 1% phiX added as control). Removal of residual genomic DNA and ribosomal RNA, preparation and sequencing of genomic libraries were performed in accordance with the manufacturer's instructions.