GSM5739112: RPE Bmal1 WT-ZT1-rep2; Mus musculus; RNA-Seq
GSM5739112
Retinal pigment epithelial cell isolation protocol was followed as previously reported (DeVera and Tosini, 2020). Briefly, eyes were enucleated under room light or dim red-light conditions (< 1 lux) and removal of the anterior segment along with the neural retina was dissected from the posterior segment that contains the RPE, choroid, and sclera. Each dissected posterior segment was placed in 50 μL of RNAlater (Qiagen, 76,104). Removal and isolation of RPE cells was done via brief sonication (Fisher, F60) on ice (< 2 s at 10% max power) and was done in ambient light conditions. Two RPE samples from one mouse were pooled into one tube. The isolated RPE cells were processed for RNA isolation with TRIzol (Ambion, 15596018) following the manufacturer's instructions. The Illumina Stranded Total RNA Prep, Ligation with Ribo-Zero Plus (Illumina, 200050529) was used for library preparation and done according to the manufacturer's instructions