Tissue preparation protocol. Animal were killed by rapid cervical dislocation. Retinas were removed immediately and placed in 1 ml of 160 U/ml Ribolock for 1 min at room temperature. Dissecting and preparing eyes for RNA extraction Retinas for RNA removed from the eye and placed in Hank’s Balanced Salt solution with RiboLock (Thermo Scientific RiboLock RNase #EO0381 40U/µl 2500U) and processed per manufacturer’s instructions (in brief form below).
- -1) Sample collection for RNA isolation.
- -2) Quickly remove the retinas with clean curved forceps after cervical dislocation of the mouse.
- -3) Put each retina in 1 ml of 160 U/ml Ribolock for 1 min in RT.
- -4) Move the retina to another tube with 50µl Ribolock, store in -80°C.
- -5) The RNA was isolated using a QiaCube and the in column DNAse procedure.
- -Quality Control: All RNA samples were checked for quality before running microarrays. The samples were analyzed using the Agilent 2100 Bioanalyzer. The RNA integrity values for each sample are presented in Table 1 below.
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