NextSeq 500 sequencing; Single cell RNA sequencing of murine gamma delta T cells from wild type and beta2 integrin-deficient (CD18 KO) lungs
Single cell RNA sequencing of murine gamma delta T cells from wild type and beta2 integrin-deficient (CD18 KO) lungs
CD18 (#003329) KO mice were purchased from The Jackson Laboratory. C57Bl/6J mice were purchased from Envigo. Lungs were perfused with 10ml 2mM EDTA in PBS prior to removal. Lung tissue was digested in 1mg/ml Collagenase D and 100ug/ml DNase I at 37 degrees C for 40 mins with shaking. γδ T cells were sorted from single cell suspensions by FACS for CD3+ TCRγδ+ cells. Single cell suspensions were loaded into a Chromium Controller (10X Genomics). cDNA was generated from polyadenylated RNA, amplified and converted to Illumina compatible libraries according to standard protocol published in Single Cell 3' Reagent Kits v3 User Guide (10X Genomics). cDNA was generated from polyadenylated RNA, amplified and converted to Illumina compatible libraries according to standard protocol published in Single Cell 3' Reagent Kits v3 User Guide (10X Genomics).