GSM6793374: SC, EU labeled, P14 [EU211]; Mus musculus; RNA-Seq
Following vitreal EU injections, we dissected out superior colliculi from three animals and isolated total RNA using TRIzol reagent (Invitrogen 15596029). Twenty µg of total RNA was reacted with biotin-azide using Click-iT™ Nascent mRNA capture kit (Invitrogen C10365) to label EU-containing RNAs with a biotin tag. The tagged RNAs were affinity-purified using MyOne™ streptavidin T1-dynabeads resuspended in 50 µl nuclease-free water. Purified RNAs were converted to double-stranded (ds) DNA using the previously described method (Shigeoka et al., 2016) with slight modification. Amplified cDNAs were converted to sequencing libraries using Nextera XT DNA library preparation kit (Illumina FC-131-1024) and sequenced in Novaseq 6000.