GSM8458003: 8D_Treated_3; Mus musculus; RNA-Seq
Mice were euthanized by cervical dislocation, and eyes enucleated and immersed in RNAlater solution. Retinas were dissected rapidly in RNAlater,and stored in RNAlater solution. Lysis was performed with the TissueLyser LT (Qiagen Gmbh, Hilden, Germany) bead mill, using one steel ball at 50 Hz for 5 min on ice. For the 5-week timepoint, total RNA was extracted using the RNeasy Mini Kit (Qiagen Gmbh, Hilden, Germany) and for the 1-week timepoint it was extracted using the RNeasy Plus Mini Kit (Qiagen Gmbh, Hilden, Germany), following the manufacturer's instructions. 500 ng of DNase I-treated total RNA was used to enrich for polyA-containing transcripts, using oligo(dT) primers bound to beads. The enriched RNA pool was fragmented and converted to cDNAs, which were ligated to sequencing adaptors containing index sequences using the Illumina TruSeq stranded mRNA sample preparation kit (Cat. #RS-122-2101, Illumina, Inc., San Diego, CA). Pools were sequenced on the Illumina HiSeq 4000 genome sequencer using 75 bp paired-end SBS chemistry.