GSM6935764: Foxp1 transgenic; Mus musculus; RNA-Seq
Cells for single cell sequencing were obtained from two P0 female littermates with the genotypes Foxp1 tg/+; Six3-Cre (Foxp1 cTG) and Foxp1 tg/+ (control). Two freshly dissected retinas from one individual animal were pooled for each sample and dissociated in PBS, 50 mM HEPES, 0.05 mg/ml DNase I and 0.025 mg/ml Liberase for 35 min with intermediate trituration. Cells were passed through a 40 μm nylon cell strainer, washed with washing buffer (1X PBS, 2% BSA, 0.1% sodium azide, 0.05% EDTA), and red blood cells were lysed in RBC lysis buffer. Cell counts were determined using a Countess. 1x 10e6 cells was resuspended in 1mL DPBS and provided to the High Throughput Genomics Core for library prep. Single cell libraries were generated with Next GEM Single Cell 3' Gene Expression Library prep v3.1 according to manufacturer's protocol (10X Genomics PN-1000128). Sequencing libraries were applied to a NovaSeq flow cell using XP chemistry workflow v1.5 (Illumina 20043131) and a 150 x 150 cycle paired end sequence run was performed using an Illumina NovaSeq 6000 instrument and S4 reagent Kit v1.5 (20028312). Libraries were sequenced to > 200 million reads per sample.