GSM9261464: R18_lung_mRNA_lane1; Mus musculus; OTHER
Cell suspensions were prepared by mechanical dissociation, followed by enyzmatic digestion using collagenase/hyaluronidase. The single cell suspension was then enriched for GFP+ cancer cells. Then, we stained each sample with it's own unique hashing antibody so that we could subsequently pool the samples for loading onto the 10x Chromium machine and later prepare sequencing libraries, and finally separate each sample in silico by it's unique hashing antibody. 3x libraries from each lane are sequenced separately on a NovaSeq machine. Transcriptome library preparation was conducted according to the Chromium Next GEM Single Cell 3' Reagents Kits v3.1 with Feature Barcoding technology kit manual. Hashtag libraries were prepared using the standard CITE-seq protocol (https://citeseq.files.wordpress.com/2019/02/cite- seq_and_hashing_protocol_190213.pdf). LARRY barcode libraries were constructed using a custom nested PCR protocol followed by indexing.