GSM5221763: B6CTRL2; Mus musculus; RNA-Seq
Tet+ and Tet- CD4 single positive thymic cells were sorted into Trizol LS.RNA was extracted using RNeasy mini kit (Qiagen) per manufacturer instructions. After ribogreen quantification and quality control by Agilent BioAnalyzer, total RNA was amplified using the SMART-seq V4 (Clonetech) ultralow input RNA kit for sequencing (12 cycles of amplification for 2–10 ng of total RNA). 10 ng of amplified cDNA was used to prepare Illumina Hiseq libraries with the Kapa DNA library preparation chemistry (Kapa Biosystems) using 8 cycles of PCR. Samples were barcoded and run on a Hiseq 4000, in a 50-bp/50-bp paired-end run, using the TruSeq SBS Kit v3 (Illumina).