GSM3619260: RNA (CD8+/GFP+) in vitro effector TCR SV40-I CD8 T cells overexpressing GFP replicate#1; Mus musculus; RNA-Seq
Samples for RNA-seq were sorted directly into Trizol LS (Invitrogen). The volume was adjusted to 1ml with PBS and samples frozen and stored at -80°C. RNA was extracted using RNeasy mini kit (Qiagen) per instructions provided by the manufacturer. After ribogreen quantification and quality control of Agilent BioAnalyzer, total RNA underwent amplification using the SMART-seq V4 (Clonetech) ultralow input RNA kit for sequencing (12 cycles of amplification for 2-10 ng of total RNA). Subsequently, 10 ng of amplified cDNA was used to prepare Illumina Hiseq libraries with the Kapa DNA library preparation chemistry (Kapa Biosystems) using 8 cycles of PCR. Samples were barcoded and run on a Hiseq 4000, in a 50bp/50bp paired-end run, using the TruSeq SBS Kit v3 (Illumina). 10 ng of amplified cDNA was used to prepare Illumina Hiseq libraries with the Kapa DNA library preparation chemistry (Kapa Biosystems) using 8 cycles of PCR. Samples were barcoded and run on a Hiseq 4000, in a 50bp/50bp paired-end run, using the TruSeq SBS Kit v3 (Illumina).
GEO Accession
GSM3619260