RNA-Seq of human ITS-2 fragments
The small RNA was first treated with T4 Polynucleotide Kinase (PNK).Then, oligonucleotide adapterswere ligated to the 5' and 3' ends of the RNA sample. First-strand cDNA synthesis was performedusing M-MLV reverse transcriptase and the 3 adapter as primer. The resulting cDNA was amplifiedwith PCR using a high fidelity DNA polymerase. 12 PCR cycles were used and  the  i5 barcode TATGCAGT as  part to the 3' sequencing adapter introduced. The cDNA was purified using theAgencourt AMPure XP kit (Beckman Coulter Genomics). The cDNA pool was single-end sequenced with a NextSeq 500 system using 150 bp read-length. Library preparation and sequencing was performed by  vertis Biotechnologie AG, Freising, Germany