GSM5290409: Bulk_RNA_CK_Baseline_rep1; Mus musculus; RNA-Seq
For bulk RNA-sequencing of sorted nuclei, samples were treated for 15 minutes with Proteinase K at 50°C and then for 13 minutes at 80°C. RNA was extracted using Direct-zol RNA MicroPrep kit according to manufacturer's instructions (cat no. R2062, Zymo Research, Irvine CA). Purified RNA samples underwent fragment analysis at the MIT BioMicro Center. Libraries were generated from samples passing quality control (DV200 < 50%). Library generation was performed using the SMARTer Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian (cat no. 634412, Takara Bio Inc), then submitted to the MIT BioMicro Center for quality control (fragment analysis and qPCR), followed by sequencing. Paired-end sequencing was performed using the Illumina NextSeq500 platform according to standard operating procedures.