GSM6615201: Spleen cell mix 2, replicate B, scRNAseq, hashtag library; Mus musculus; RNA-Seq
Spleens were mechanically dissociated and then filtered through a 70μm cell strainer to isolate a single cell suspension. Tumors were mechanically dissociated in a cocktail containing RPMI, DNase1, and collagenase D using gentlemacs C tubes, followed by incubation at 37C in an incubator for 30 minutes and then filtering the cell suspension through a 70μm cell strainer. Isolated single cell suspensions were then stained with hashtag antibodies and cell surface antibodies for FACS. Isolated Live, CD45+, CD11b+, CD11c+ cell were used as the input for the 10X genomics chromium platform using a 5' sequencing kit and protocol. Library construction was performed according to manufacturers instructions (10X genomics, 5' kit version 2)