GSM8972219: GEX + HTO (Sample tags: 7, 8, 9, 10, 11, 12); Mus musculus; RNA-Seq
B16.F10 tumors were isolated on day 21 after inoculation. Excised tumors were cut into small pieces (2-4 mm), transferred into gentleMACS™ C tubes (Miltenyi Biotec; 130-093-237) and digested with the Tumor Dissocation Kit, mouse (Miltenyi Biotec; 130-096-730) according to the manufacturer's instructions using the gentleMACS™ Octo Dissociator with Heaters (Miltenyi Biotec; 130-096-427). CD8+ and CD4+ tumor-infiltrating lymphocytes were isolated using CD4/CD8 (TIL) (Miltenyi Biotec; 130-116-480)  Microbeads according to the manufacturer's instructions using magnetic separation (magnetic activated cell sorting (MACS)). Single cell capturing, isolation of RNA and synthesis of cDNA was performed using the BD Rhapsody Single-Cell Analysis System (BD Biosciences) according to manufacturer's instructions (All reagents required for this purpose are listed in document BD Biosciences, Doc ID: 210966 Rev1.0). Each sample was tagged with a special sample tag in order to allow multiplexing of samples on the same cartridge. Sequence of sample tags can be foundin the BD document Doc ID 210970. Whole transcriptome libraries and sample tag libraries were created using BD WTA Amplification Kit (BD Biosciences,633801) with random primers followed by amplification and insertion of sequencing adaptors following the BD Rhapsody System mRNA WTA and Sample Tag Library Preparation Protocol (BD Biosciences, Doc ID: 23-24119 Rev1.0). Quality and quantity of cDNA libraries were analyzed on a Bioanalyzer2100 (Agilent). Sequencing (paired end reads, PE150) was performed on a NovaSeq 6000 (Illumina) at Novogene (Cambridge; UK). For processing of the sequencing raw data BD Rhapsody WTA analysis pipeline version 2.0 was used.
GEO
GSM8972219_r1