GSM7715053: SAM24380130_VG_WT_2; Mus musculus; RNA-Seq
Due to the low number of nodose neurons we pooled VG from 3 mice for nuclei isolation per analyzed sample (4 samples total). Harvested VG were incubated in 0.5 mL Lysis buffer (10 mM NaCl; 10 mM Tris pH 7.5; 3 mM MgCl2; 0.2 U/mL RNase inhibitor) for 5 min on ice. VG were then careful ly homogenized using a dounce homogenizer and spun at 300 g for 5 minutes. Supernatant was discarded and the pellet was resuspended in 0.5 mL PBS+ 3% BSA+ 0.2 U/mL RNase inhibitor containing 5 uL RNase free DNaseI and incubated at RT for 2 minutes. The solution containing nuclei was gently triturated, pelleted and re suspended in PBS+ 3% BSA+ 0.2 U/mL RNase inhibitor. Isolated nuclei were injected into Chromium for GEM generation and subsequent library construction (V3 of t he Chromium 3' EXP reagent kit; 10x Genomics). cDNA of the first RT reaction as well as the final library was QC'ed via High Sensitivity DNA chip bioanalyzer ( Agilent). 10x Genomics 3' v3 chemistry