GSM5460405: Bispecific_T24; Mus musculus; RNA-Seq
MC38 tumors were mechanically dissociated by using the gentleMACS OCTO Dissociator and then filtered over 70uM nylon mesh filter. Spleens were treated for 25 minutes at 37 degrees with 25 ug Liberase TL (Roche) then mechanically dissociated by processing over a 70uM nylon mesh filter before treatment with ACK Lysis Buffer (Gibco) at room temperature for 3 minutes. Single cell suspension of the samples were then enriched for CD45+ cells using the Miltenyi mouse CD45(TIL) Microbeads Kit (Miltenyi BioTec) according to manufacturer's instructions for the LS columns. Single-cell emulsions were obtained using the 10X Genomics Controller and the v2 library and Gel Bead kit (10x Genomics). RNA-sequencing libraries were prepared as instructed by the 10x 3' v2 kit protocol. Resulting libraries were sequenced on an Illumina NextSeq using a NextSeq 500/550 v2.5 High Output Kit.