title: GSM6289763: CD4, replicate1, scRNAseq_HTO; Mus musculus; RNA-Seq
library_construction_protocol: Spleens were flushed with PBS, passed through a cell strainer, and lysed with red blood cell lysis buffer. Small intestines were excised, sectioned and incubated in RPMI containing 5mM EDTA, 1mM DTT and 3% v/v calf serum at 37°C for 20 minutes. The resulting digest was passed through a cell strainer and IELs isolated by centrifugation over a Histopaque gradient (Sigma). For LPL isolation, tissues were minced and digested with Liberase TL (Roche) (2.5mg/ml) and DNAse (Sigma) (0.02mg/ml) at 37°C for 30 minutes and passed through a cell strainer. Colons were processed the same as SI-LPLs without an IEL separation. For skin lymphocyte isolation, mouse ears were digested in HBSS CollagenaseIV (1mg/ml) at 37°C for 2 hours and the resulting digest  was passed through a cell strainer. Final cell suspensions were in FACS buffer containg 3% v/v calf serum and  2mM EDTA. Library was performed according to the manufacter's instructions (single cell 3' v3 protocol, 10x Genomics).