GSM3554184: Spleen_CD69neg_3; Mus musculus; RNA-Seq
Total RNA was purified with miRNeasy Micro Kit (Qiagen) from FACS sorted LCMV-specific CD4 memory T cells isolated from spleen and bone marrow. RNA quality was determined on a Bioanalyzer using the RNA 6000 Pico Kit (Agilent). For poly-A dependent cDNA synthesis and a first amplification step the Smart-Seq v4 mRNA Ultra Low Input RNA Kit (Clontech) was used. After quality control (High sensitivity DNA Kit, Agilent) and concentration measurement (Qubit dsDNA HS Assay Kit, Invitrogen), 150 pg of the purified cDNA was used for tagmentation and library completion with the Nextera XT library preparation kit (Illumina). In the following, 2x75 nucleotides paired-end sequencing was performed on a NextSeq 500 using a mid-output flow cell.
GEO Accession
GSM3554184