GSM5218917: MibrT2 Treg co-cultured 2; Mus musculus; RNA-Seq
Frozen cell pellet samples were disrupted in Trizol®. Chloroform was added to the reaction and mixed vigorously. Following phase separation by centrifuging for 15 minutes at 12,000g, the aqueous phase was removed to a clean microcentrifuge tube. RNA was precipitated by isopropanol and pelleted by centrifugation for 10 minutes at 12,000g. The SMART-Seq HT Kit was used to generate cDNA from total RNA The sequencing library was constructed by following the Illumina Nextera XT Sample Preparation Guide. One nanogram of input cDNA was tagmented (tagged and fragmented) and amplified using the Illumina Nextrera XT kit. Sequence libraries of each sample were finally equimolarly pooled and sequenced on an Illumina Nextseq 500 system, using a paired-end 75-bp strategy.