HD1   FL33RNAseq
PBMCs were obtained from healthy donors or indicated patients with written informed consents. Normal T cells sorted from PBMCs sample, using EasySepTM Human T Cell Enrichment Kit. The fragmented mRNA was used as a template and random oligonucleotides were used as primers to synthesize the first strand of cDNA in the M-MuLV reverse transcriptase system, followed by degradation of the RNA strand with RNaseH and synthesis of the second strand of cDNA with dNTPs under the DNA polymerase I system. The purified double-stranded cDNAs are end-repaired, A-tailed and ligated to sequencing connectors. cDNAs of about 250-300bp are screened with AMPure XP beads, PCR amplified and PCR products are purified again with AMPure XP beads to obtain the final libraries. The kit used for library construction is NEBNext Ultra RNA Library Prep Kit for Illumina.