GSM8967582: Treg_Sp_IL2_JES6_1h_2; Mus musculus; RNA-Seq
For each sample, 1000 cells were sorted directly into the lysis buffer [5uL TCL buffer (Qiagen) supplemented with 1% 2-mercaptoethanol (Sigma)]. Total RNA was purified using RNAClean XP beads (Beckman Coulter). Polyadenylated mRNA was selected using an anchored oligo(dT) primer (5′–AAGCAGTGGTATCAACGCAGAGTACT30VN-3) and then reverse transcribed to cDNA. First-strand cDNA was subjected to limited PCR amplification followed by Tn5 transposon-based fragmentation using the Nextera XT DNA Library Preparation Kit (Illumina). Libraries were constructed following the Smart-seq2 RNA-seq library preparation protocol.