QuantSeq 3'mRNA-Seq of bonemarrow tissue of mus musculus upon Toxoplasma gondii infection 0dpi
Bone marrow samples of uninfected (d0) and T. gondii-infected (ME49, bradyzoites from 10 cysts, i.p.) WT and LTR /- mice were generated via centrifugation. Briefly, murine femora were cut open at joint-side and placed in a punctured 0.5 ml tube that were then placed in a 1.5 ml tube. After centrifugation (12,100 x g, 15 sec) and red blood cell (RBC) lysis with Erylysis-buffer (Morphisto, Germany), remaining cells were stabilized in RNAprotect Tissue Reagent (Qiagen, Germany) according to the manufacturers protocol. Total RNA was isolated using the RNeasy Plus Mini Kit (Qiagen, Germany) according to the manufacturers protocol. RNA quality was analyzed and 25ng of total RNA per sample were used as input for the QuantSeq 3 mRNA-Seq V2 Library Prep Kit with UDI (Lexogen, Austria), which was used to generate the sequencing library according to the manufacturers protocol.