title: GSM4306703: GM7810S2; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA from primary fibroblasts was extracted using the RNeasy Mini Kit (Qiagen), including DNAse I treatment to remove genomic DNA contamination. Material was quantified using RiboGreen (Invitrogen) on the FLUOstar OPTIMA plate reader (BMG Labtech) and the size profile and integrity analysed on the 4200 TapeStation (Agilent, RNA ScreenTape). Input material was normalised to 50 ng prior to library preparation. 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 normalised 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 two lanes (669382 and 670822) of a HiSeq4000 75bp platform (Illumina, HiSeq 3000/4000 PE Cluster Kit and 150 cycle SBS Kit).
attributes.tag: GEO Accession
attributes.value: GSM4306703