title: GSM6132493: Mm_AMB_N176; Mus musculus; RNA-Seq
library_construction_protocol: On the next day, nAmb tissue was homogenized and purified by density-gradient centrifugation into a single-nuclei suspension as previously described (Habib et al., 2016; Todd et al., 2020). The single-nuclei suspension was sorted by FACS to isolate one H2b-mCherry+ nucleus per well of three 96-well plates, which were then centrifuged at 2,500rcf, frozen on dry ice, and stored at -80°C until use. After purifying the RNA by RNA-clean SPRI reagents (1.5x ratio of SPRI to sample), cDNA libraries were generated from polyadenylated mRNA of each sample using Smart-Seq2 (Picelli et al., 2014), modified as previously described (Tao et al., 2021). Illumina sequencing libraries were prepared from single-nuclei cDNA samples as described previously (Tao et al., 2021) and then sequenced by Illumina Next-Seq 500.