GSM8559649: P75 Control-3; Mus musculus; RNA-Seq
A total of 50,000 FACS-purified cells were collected directly into the RLT Plus buffer (QIAGEN, 1053393), and RNA was purified from these sorted cells with the RNeasy Plus Micro Kit (QIAGEN, 74034) and DNase1 treatment (QIAGEN, 79254) according to the manufacturer's instructions. For RNA-seq library preparation, sample quality was measured using an Agilent Bioanalyzer. Only samples with RNA integrity numbers of >8 were used for library preparation. For generating RNA-seq libraries, 50 ng of total RNA was subjected to polyadenylate selection using Universal Plus mRNA-Seq with NuQuant (TECAN, 0520-A01). Following elution, mRNA was subjected to fragmentation at 94°C for 8 min. First strand, second-strand cDNA synthesis, end repair, adapter ligation, and amplification were carried out by following the manufacturer's instructions. Libraries were sequenced on the Illumina Hiseq Platform.