title: GSM4495686: Abdo DFAT tet-shCON_dox_3; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA purification and on-column DNaseI-treatment were performed using the RNeasy Mini kit (Qiagen). Library preparation, cDNA sequencing and analysis were performed at the Oxford Genomics Centre (Wellcome Trust Centre for Human Genetics, Oxford, UK). Polyadenylated transcript enrichment and strand specific library preparation was completed using NEBNext Ultra II mRNA kit (NEB) following manufacturer's instructions. Libraries were amplified on a Tetrad (Bio-Rad) using in-house unique dual indexing primers (based on DOI: 10.1186/1472-6750-13-104). Individual libraries were normalized using Qubit, and the size profile was analysed on the 2200 or 4200 TapeStation. Individual libraries were normalised and pooled together accordingly. The pooled library was diluted to ~10 nM for storage. The 10 nM library was denatured and further diluted prior to loading on the sequencer. Paired end sequencing was performed using a HiSeq4000 75bp platform (Illumina, HiSeq 3000/4000 PE Cluster Kit and 150 cycle SBS Kit), generating a raw read count of 20-25 million reads per sample.
identifiers.namespace: GEO
identifiers.id: GSM4495686