GSM8991950: Active_Treg_WT_rep3; Mus musculus; RNA-Seq
RNA extraction was  performed according to the manufacturer's instructions. Briefly, phase separation in cells lysed in 1mL TRIzol Reagent was induced with 200 µL chloroform. RNA was extracted from 350 µL of the aqueous phase using the miRNeasy Micro Kit (Qiagen catalog # 217084) on the QIAcube Connect (Qiagen) according to the manufacturer's protocol. Samples were eluted in 15-18 µL RNase-free water. After RiboGreen quantification and quality control by Agilent BioAnalyzer, 1-2 ng total RNA with RNA integrity numbers ranging from 5.5 to 9.3 underwent amplification using the SMART-Seq v4 Ultra Low Input RNA Kit (Clonetech catalog # 63488), with 12 cycles of amplification. 4.5-8 ng of amplified cDNA was used to prepare libraries with the KAPA Hyper Prep Kit (Roche 07962363001) using 8 cycles of PCR.