GSM8285573: MT002A: T1 - Batch 1; Homo sapiens; RNA-Seq
UMI-bc containing lentiviral constructs were transduced into Hs766T cell line with MOI <0.1 followed by puromycin selection. After all the non-transduced cells were dead, we seeded 8000 cells / well (multiple wells in total), followed by incubation which lasted approx. 1-2 population doublings (daughter cells were created for each UMI-BC containing cell). At this point we performed the 1st 10x chromium run (time point 1) so that 50% of the cells were used for this this initial time point run, and the remaining 50% of the cells were cultured further for the 2nd time point. The 2nd time point chromium run was done approx. 10 population doublings later. Portion of amplified cDNA-library was re-amplified with specific ORF_BC_amplif_oligo_F & ORF_BC_amplif_R to enrich for the lineage tracing UMI-BC (the same oligos were used in the Pooled TF overexpression assay in our publication). Finally this amplified fragment was spiked in to the whole-transcriptome part of the library with estimated 10% abundance. scRNA-seq based bar-coded lineage tracing