GSM1541959: MethylC-seq_excitatory_neurons_rep2; Mus musculus; Bisulfite-Seq
The neocortex was rapidly dissected from 1-2 adult mice on ice, and the INTACT method was used to affinity purify GFP+ nuclei. Isolated nuclei were resuspended in PBS for DNA purification using the DNeasy Blood and Tissue kit (Qiagen 69504) following the standard kit protocol. MethylC-seq libraries were constructed as previously described [Lister R. et al. Global epigenomic reconfiguration during mammalian brain development. Science 341, 1237905 (2013)], except that samples were PCR amplified with KAPA HiFi HotStart Uracil+ ReadyMix (Kapa Biosystems KK2802) using the following PCR conditions: 2 minutes at 95°C, 4 cycles of [15 seconds at 98°C, 30 seconds at 60°C, 4 minutes at 72°C], and 10 minutes at 72°C. Libraries were sequenced on an Illumina HiSeq 2000 up to 101 cycles.