title: GSM8368830: LOS_E6_S228_Aligned.quant; Mus musculus; RNA-Seq
library_construction_protocol: Cells from 3-6 animals were pooled prior to FACS-isolation. In brief, cells were sorted into hypotonic lysis buffer, snap frozen in liquid nitrogen and stored at -80oC until all samples were collected. Cells were lysed by heating at 72oC for 3 min, followed by reverse transcription of mRNA using dT30 oligos, template switching oligos and Maxima H- reverse transcriptase. The whole transcriptome was amplified (15 cycles) by KAPA HiFi DNA polymerase (Roche), and then size-selected using 0.6X AmpPure XP beads (Beckman Coulter). To exclude cells with poor amplification, and wells containing multiple cells, qPCR for Gapdh was performed. Illumina sequencing libraries were indexed with unique 5' and 3' barcode combinations (up to 384 cells) using the Nextera XT DNA library preparation kit (Illumina). Libraries were pooled and size-selected with 0.9X AmpPure XP beads. Prior to sequencing on Illumina NextSeq500 using a 75 bp paired-end read mid-output setting, library quality was assessed by TapeStation (Agilent).