NextSeq 500 paired end sequencing; RNA sequencing of primary isolated unstimulated murine splenic DC1 and DC2 from specific pathogen free (SPF), Germ free (GF) and Interferon alpha/beta receptor 1 (Ifnar) knock out mice
RNA sequencing of primary isolated unstimulated murine splenic DC1 and DC2 from specific pathogen free (SPF), Germ free (GF) and Interferon alpha/beta receptor 1 (Ifnar) knock out mice
Cell were sorted and lysed in Trizol. Total RNA was isolated from Trizol in combination with the miRNeasy Micro Kit (Qiagen). For poly-A-dependent cDNA synthesis and a first amplification step 10 ng of total RNA was used as input in the Smart-Seq v4 mRNA Ultra Low Input RNA Kit (Clontech) and processed according to the manufacturer's instructions. 1 ng of the purified cDNA was used for tagmentation and library completion with the Nextera XT library preparation kit (Illumina).
Experimental Factor: strain
specific pathogen free C56BL/6
Experimental Factor: genotype
wild type genotype
Experimental Factor: cell type
CD103-positive dendritic cell