Single-cell RNA-Seq of Mus musculus : PN4 Posterior cerebellum control
Single-cell RNA-Sequencing was carried out between PN4 control and Tlx3 cKO posterior cerebellum. Posterior cerebellar tissue was freshly dissociated using Papain solution in HBSS containing L-Cysteine, and papain was inactivated using ovomucoid solution. Following this, samples were centrifuged at 1800 rpm for five minutes. Supernatant were carefully removed and cells was resuspended in 1ml of 10mg/ml BSA. After another round of centrifugation at 1800 rpm for five minutes, cells were resuspended in 1ml 1X PBS. Cell death was analyzed using trypan blue and cells were counted using a haemocytometer. Approximately 1000-2000 cells of control and Tlx3 cKO single-cell suspensions were loaded onto a 10X chromium platform for generating single-cell Gel Bead-In-EMulsion (GEMs). Libraries were generated with Chromium Next GEM Single Cell 3 LT Reagent Kits v3.1(Dual index) as per 10X genomics protocol and were sequenced.The sequencing of libraries was performed on an Illumina Novaseq platform.