GSM5174168: 4967_5_WT_Treg_Lymph_nodes_-_8_5_18_ACAGTGA; Mus musculus; RNA-Seq
Tregs were flow cytometrically sorted in RLT buffer (qiagen) +1% b-mercapto-ethanol for the subsequent extraction of total RNA using RNAeasy mini kit (Qiagen). Total RNA was extracted using according to the manufacturer's protocol. Library preparation was performed on RNA samples with a measured RNA Integrity Number (RIN) between 8.0 and 10.0 as assessed by the 2100 Bioanalyzer using a Nano chip (Agilent). Strand-specific libraries were generated using the TruSeq Stranded mRNA sample preparation kit (Illumina Inc.) according to manufacturer's instructions (Illumina, Part # 15031047 Rev. E). RNA libraries were prepared using standard Illumina protocols, analyzed for size and quantity of cDNAs on a 2100 Bioanalyzer using a DNA 7500 chip (Agilent), diluted and pooled in multiplex sequencing pools. The libraries were sequenced as 65 base single reads on a HiSeq2500 (Illumina).