GSM7061446: IL33-treated LTBR-positive myeloid cells; Mus musculus; RNA-Seq
Fluorescence activated cell sorting-purified lymhotoxin-beta-positive and -negative myeloid cells were loaded onto to the 10x Genomics Chromium platform to generate Gel Beads-in-Emulsion (GEM). Samples were incubated at 53 °C for 45 min in a C1000 Touch Thermal cycler to generate polyA cDNA barcoded at the 5'end. GEMs were broken and single-strand cDNA was cleaned up with DynaBeads MyOne Silane Beads. The cDNA was amplified for 16 cycles. 50ng of cDNA was enzymatically fragmented, end repaired, A-tailed, subjected to a double-sided size selection with SPRIselect beads and ligated to adaptors provided in the kit. Single-cell RNA-seq libraries were prepared according to 10X Genomics specifications (Chromium Single Cell V(D)J User Guide PN-1000006), see above.