GSM4725813: RPMC isolated from specific-pathogen free housed mice [RPMC_SPF_3]; Mus musculus; RNA-Seq
Cells were directly FACS-sorted into RLT+ lysis buffer (Qiagen), containing 10 µl/ml β-mercaptoethanol (Roth). Total RNA was extracted with the RNeasy Plus Micro kit (Qiagen) according to manufacturer's instructions. cDNA conversion was carried out using the SMART‑Seq v4 Ultra Low Input RNA Kit (Takara Clontech Laboratories) according to manufacturer's instructions, followed by PCR product purification with Agencourt AMPure XP magnetic beads (Beckman Coulter). Libraries were prepared with the Nextera XT DNA Library Prep Kit (Illumina) employing enzymatic tagmentation and simultaneous adapter ligation before subsequent indexing and fragment amplification. PCR product clean-up and size selection was performed with the Agencourt AMPure XP magnetic beads. RNA and cDNA quality and content, as well as the correct sizing of cDNA library fragments was verified using the Agilent Technologies 2100 Bioanalyzer profiles and Qubit measurements after each step.  Libraries were pooled and sequenced at the genome analytics facility of the Helmholtz Centre for Infection Research on Illumina HiSeq2500 using 50 bp single-end reads.