GSM3674849: ORGN-Lgr5-plate14-labeled; Mus musculus; OTHER
After incubation with EU, cells were dissociated, fixed with paraformaldehyde, and azide-biotin was clicked in situ after cell permeabilization to mark EU-labeled RNA. We then sorted single cells in 384-well plates and performed first strand synthesis with poly-T primers containing a cell barcode, a unique molecular identifier, a 5' sequencing adapter, and the T7 promoter. Cells were pooled and streptavidin magnetic beads were used to separate EU labeled and unlabeled mRNA/cDNA hybrids. Library preparation was performed for both fractions separately. Runs contain the 6-nucleotide long UMI in the first nucleotides of Read 1, followed by the 8-nucleotide long cell-specific barcode. Illumina TruSeq adapaters
GEO Accession
GSM3674849