Microarray analysis of circRNAs

Microarray analysis is widely used to identify altered circRNAs in in vivo studies ²¹. Three pairs of retinas in each group were used for the analysis. Sample labeling and microarray hybridization were conducted according to the standard protocols Arraystar (Rockville, MD, USA). Briefly, total RNAs were digested with Rnase R (Epicentre, Madison, WI, USA). A random priming method with the Arraystar Super RNA Labeling Kit was utilized to amplify the enriched circRNAs and to transcribe the circRNAs into fluorescent cRNA. Labeled cRNAs were hybridized onto Mouse circRNA Array V2 (8x15K, Arraystar). Then the slides were washed, and the arrays were then scanned by Agilent Scanner G2505C, and the images were analyzed by an Agilent Feature Extraction software (version 11.0.1.1). Detected circRNAs were regarded as significantly differentially expressed by the value of fold change≥1.5 and P<0.05.