title: GSM9223238: T cells, PERK-KO, Allo stimulation-3; Mus musculus; RNA-Seq
library_construction_protocol: RNA was harvested using TRIZOL (invitrogen). 0.5ug of total RNA was used to construct sequence libraries. First, the mRNA is fragmented randomly by adding fragmentation buffer, then the cDNA is synthesized by using mRNA template and random hexamers primer, after which a custom second-strand synthesis buffer (Illumina) , dNTPs, RNase H and DNA polymerase I are added to initiate the second-strand synthesis. Second, after a series of terminal repair, A ligation and sequencing adaptor ligation, the double-stranded cDNA library is completed through size selection and PCR enrichment.