GSM5237891: Sample 52_TENA2517_P16_R7; Mus musculus; RNA-Seq
FACS-sorted cells were collected as 5-cell pools in 2.5µl of 0.2% TX-100 and 2U/μl RNase inhibitor (TakaraBio, 2313B) in PBS into 96-well plates (Thermo Fisher Scientific, 4346906) and kept in -80°C prior to use. RNAseq libraries were prepared according to the Smart-seq2 method (Picelli, 2014). 5-cell RNA-seq: RNA/cDNA preamplification and Nextera XT library preparation reactions were set up with a Mosquito HV system (SPT Labtech). After reverse transcription and amplification, the concentration of cDNA generated from each sample was measured using PicoGreen (Thermo Fisher Scientific). cDNA was normalized to 0.1-0.15ng/µl and 0.75 µl were used for library preparation with the Nextera XT DNA library preparation kit (Illumina). Multiplexed library pools were then pooled and PE 2x51 sequenced with an Illumina NovaSeq 6000 system, v1 reagents.