GSM4634764: Neuronal nuclei Col_rep1; Mus musculus; RNA-Seq
Nuclei were isolated using the NucleiEZ lysis buffer with modifications, as described in May-Zhang et al. FACS was performed to isolate neuronal nuclei and glia, which were gated for 7AAD+ and high intensity of CFP from the Phox2b-CFP reporter and collected into a 1.5-mL RNAse-free tube filled with 350 uL of RNA lysis buffer RLT from the RNeasy Micro kit. cDNA libraries were prepared using the Takara SMARTer kit according to llumina's instructions. Libraries were sequenced using the NovaSeq as 1x100bp single-end reads. Each sample had a mean depth of approximately 138 million reads and a mean alignment rate of 99.01%.