Th17
The purified mRNA was fragmented at 90C in the presence of divalent cations. Then using Superscript II Reverse Transcriptase (Life Technologies), fragments were converted into cDAN using random hexamers and. Second strand cDNA was prepared on the first strand template using RNAase and DNA polymerase I. The cDNAs so obtained were cleaned using Beckman Coulter SPRI beads. Then proceed for  sequencing libraries preparation using TruSeq RNA Library Prep Kits for Illumina (NEB, USA) according to the manufacturers instructions, and index codes were added to attribute sequences to each sample.