GSM2302811: WT_R2; Mus musculus; RNA-Seq
Retinas were dissected in cold sterile HBSS with Ca and Mg (Gibco) and stored at -80 ⁰C. Total RNA was extracted with TRIzol (Invitrogen) and purified with RNeasy Mini Kit (Qiagen) with on-column DNaseI digestion (Qiagen). Polyadenylated mRNA was selected from total RNA using Dynabeads (Invitrogen). The mRNA was fragmented and reverse-transcribed to double-stranded cDNA using random hexamers, blunt-ended, 3’-adenylated, and ligated to sequencing adapters. Ligated fragments were amplified for 12 cycles with primers to incorporate unique sample barcodes. Samples were pooled and subjected to 1x50bp sequencing on one lane of Illumina HiSeq 2000.