GSM8191511: C57BL/6J, Young mouse4, RPE/choroid, 2-3 months; Mus musculus; RNA-Seq
All mice were humanely euthanized by administering CO2 gas followed by cervical dislocation. The eyes were immediately enucleated to collect the neural retina and RPE/choroid tissues. The isolated eyes were placed in the cold petri dish under a surgical microscope. With careful precision, the anterior parts, including the cornea, lens, and iris, as well as connective tissues, muscles, and optic nerve, were excised. Radial cuts were made symmetrically in a four-leaf pattern, allowing for the careful removal of the neural retina from the mouse eyecup. The pigmented RPE/choroid layer was gently scraped from the sclera. RPE/choroid and neural retina tissues were immediately transferred to TRIzol reagent (Invitrogen 15596026, Carlsbad, CA). Subsequently, RNA purification and on-column genomic DNA digestion were conducted using the Pure Link RNA Micro Kit (Invitrogen 12183016, Carlsbad, CA). Ribosomal RNAs were removed with the Qiagen FastSelect -rRNA HMR Kit (Human/Mouse/Rat). RNA libraries were constructed using NEXTFLEX Rapid Directional RNA-Seq Kit 2.0 (Perkin Elmer, Waltham, MA) as per manufacturer's instructions.