GSM7401100: mouse 2, replicate 1, scRNAseq, TCR; Mus musculus; RNA-Seq
5 x 10^6 magnetic column purified CD4 T cells harvested from spleens of two B6.H2g7 donor mice were adoptively transferred to two separate NOD.Rag1^null mice and recovered from host spleens 14 days later by flow cytometric sorting for single cell sequencing analysis. Splenocytes from NOD.Rag1^null mice were individually homogenized using glass homogenizers in ice cold HBSS. Cells were washed and resuspended in PBS after which cells were incubated with antibodies against CD4 and T cell receptor beta and a Viability dye. Cells were washed and resuspended in PBS supplemented with 0.04% w/v BSA and 0.4 U/ul RNase I. Alive CD4 T cells were sorted into the same buffer using a BDFACS Melody (BD Bioscience). Gene expression library and TCR library was performed according to the manufacter's instructions (single cell 5' v2 protocol, 10x Genomics)