GSM8422056: 35_D1_Ad26.MVA; Homo sapiens; RNA-Seq
Venous peripheral blood (up to 3ml) was collected directly in TempusTM RNA tube immediately pre-vaccination, then 10 days post-first vaccine and 7 days post-second vaccine. RNA was extractedusing theTempus™spin RNAisolationkit(Invitrogen, USA).Eluted RNA was quantifiedusingQubit™ RNA BR Assay Kit(Invitrogen, USA) and quality control was conducted usingthe Agilent RNA ScreenTape assay on the 4200 TapeStation System (Agilent, USA). RNA was ribodepleted and globin depleted usingRibo-ZeroTM Gold rRNA removal Kit (Illumina, USA). RNA was converted to cDNA, second-strand cDNA synthesis incorporated dUTP. The cDNA was end-repaired, A-tailed and adapter-ligated and prior to amplification, samples underwent uridine digestion. The prepared libraries were size selected, multiplexed and quality controlled before 75bp paired-end sequencing (HiSeq4000). Sequencing was conducted at the Wellcome Trust Centre for Human Genetics (Oxford, UK).