GSM2574902: GTT13_P; Mus musculus; RNA-Seq
Equal numbers of Tregs (280 cells) were sorted from the thymus and placenta of E17.5 pregnant Bl6 females, mated with syngeneic Bl6 males. To increase robustness, the 280 placental Tregs were purified from 5 individual placentas per pregnant female (56 Tregs/placenta). Cells were sorted directly into a cell lysis buffer consisting of a 0.2% (vol/vol) Triton X-100 solution and an RNase inhibitor. For the generation of full-length cDNA, the Smart-seq2 protocol (Picelli, S. et al, Nat Protoc. 2014 Jan;9(1):171-81. doi:10.1038/nprot.2014.006) was used. The subsequent library preparation from the amplified cDNA was performed using the Nextera XT DNA library prep kit (Illumina). For sequencing, libraries were pooled, diluted and sequenced on Illumina HiSeq 3000 using 50 bp single-read chemistry.