GSM2124589: FD6; Gallus gallus; RNA-Seq
Total RNA was isolated from the retina/RPE/choroid using the miRNeasy Mini Kit (Qiagen) including DNase digestion. RNA quality and quantity was assessed on the 2100 Bioanalyzer. All samples had an RNA integrity number (RIN) of >8. RNA quantity was also assessed on the Qubit 2.0 Fluorometer. Using an average of concentration measures obtained from Qubit and Bioanalyzer assays, 2.5μg of RNA from each sample was used for library preparation. Libraries were prepared using the TruSeq Stranded mRNA LS kit (Illumina) with dual indexing according to the manufacturer’s instructions. The generated libraries were assessed on the 2100 Bioanalyzer to ensure an average size distribution of approximately 280bps, then quantified on the Qubit 2.0 Fluorometer and by qPCR. Libraries were normalised to 10nM in Tris-HCl, pooled, and prepared for cluster generation on the Illumina cBot using the TruSeq SR Cluster Kit V3-cBot (Illumina) with denatured template DNA diluted to 7pM. The flow cell and sequencing reagents (TruSeq SBS Kit V3; Illumina) were loaded on the Illumina HiSeq 1500 and a dual-index, single-end, 100bp sequencing run performed.
GEO Accession
GSM2124589