title: GSM6436078: KI_DN3_TCR; Mus musculus; RNA-Seq
library_construction_protocol: DN cells (CD4-CD8-) were purified by negative selection (magnetic beads) from the thymus of  C57BL/6J and GSK3-KI mice with the following antibodies: anti-MHC class II (M5/114/15/2), anti-CD11b (M1/70), anti-CD4 (GK1.5), anti-CD8 (Tib105), and anti-kappa (187.1). B220- T cells; anti-MHC class II (M5/114/15/2), anti-CD11b (M1/70), and anti-B220 (RA3-6B2). Then DN Thymocyte subsets DN3 (CD44-CD25+)  and DN4 (CD44-CD25-)  were sorted out (BD FACS AriaIII high-speed cell sorter) with anti-CD4, anti-CD8,  anti-CD44, and antiCD25 (BioLegend Way, San Diego, CA). 5' end single cell gene expression and VDJ libraries were prepared following manufacturers guidelines (10x Genomics)