GSM6430031: Small intestine_YP147_2; Mus musculus; RNA-Seq
Dendritic cells were FACS sorted into RLT+ lysis buffer (Qiagen) which contained 10 µl/ml β-mercaptoethanol (Roth). In the following, the total RNA was extracted with the RNeasy Plus Micro kit (Qiagen) according to manufacturer`s instructions. RNA libcDNA 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.raries for RNA-seq were prepared using SMARTER mRNA-Seq Library Prep Kit following manufacturer's protocols. Sequencing of libraries was carried out at the sequencing facility on Illumina NovaSeq6000 using 50 bp single-end reads and quality of sequenced libraries was verified with FastQC.