title: GSM5098818: Colon26 tumors, CD45+ cells, treated (hashtag library); Mus musculus; RNA-Seq
library_construction_protocol: Tumor were cut into small pieces with a razor blade and digested enzymatically using 0.25% Liberase TM (Merck) and 20 kU/mL DNase I (Sigma-Aldrich, St Louis, MO) for 60 min at 37°C. Tumor cell and dead cell were removed by 40% Percoll (GE Healthcare, Buckinghamshire, UK) density separation. For scRNA-seq, cDNA libraries and hashtag libraries were prepared combined by an BD Rhapsody system and an optimized process similar to our magnetic bead-based bulk RNA-seq method (Shichino et al. JCI insight 2019). Sequencing libraries were generated by using NEBNext UltraII FS library prep kit (New England Biolabs), and QC of cDNA/Hashtag libraries was performed by MultiNA (Shimazu) and qPCR library quantification assay (KAPA). BD Rhapsody WTA