GSM5082183: RV05_N2: HIF1_AAA_donor_B; Mus musculus; RNA-Seq
On day 5 post-tranduction OT-I cells were stained with Near-IR Dead Cell Stain Kit (Thermo Fisher) followed by surface staining of CD8a and Thy-1.1 and sorted on Thy-1.1 surface expression on a BD FACSAria Fusion cell sorter (BD Biosciences) directly into RLT Plus lysis buffer (Qiagen) To construct libraries suitable for Illumina sequencing the Illumina TruSeq Stranded mRNA Sample preparation protocol which includes cDNA synthesis, ligation of adapters and amplification of indexed libraries was used. The yield and quality of the amplified libraries were analysed using Qubit by Thermo Fisher and the Agilent Tapestation. The indexed cDNA libraries were normalised and combined and the pools were sequenced on the Nextseq 550 for a 50-cycle v2.5 sequencing run generating 75 bp single-end reads. Basecalling and demultiplexing was performed using CASAVA software with default settings generating Fastq files for further downstream mapping and analysis.
GEO Accession
GSM5082183