Human circrna array v2 8x15k

Microarray analysis of circRNA expression was performed on a subset of 20 samples—12 patients (6 females, 6 males) and 8 age- and sex-matched controls. Total RNA from each sample was prepared for the microarray analysis according to the manufacturer’s protocol (Arraystar, USA). Briefly, total RNA was digested with RNase R (Epicentre, Inc., USA) to enrich circular RNAs. Enriched circular RNAs were amplified and transcribed into fluorescent complimentary RNA utilizing a random priming method (Arraystar Super RNA Labeling Kit; Arraystar, USA) and then hybridized onto the Arraystar Human circRNA Array V2 (8x15K, Arraystar, USA). Slides were washed and the arrays were afterwards scanned by the Agilent Scanner G2505C.
Acquired array images were analyzed using Agilent Feature Extraction software (version 11.0.1.1). Quantile normalization and subsequent data processing were performed using the R software limma package. Differentially expressed circRNAs with statistical significance between two groups were identified through volcano plot filtering. Fold change filtering was used to identify differentially expressed circRNAs between two samples. Distinguishable circRNA expression patterns among samples were identified through hierarchical clustering.

We have completed the Arraystar Human circRNA Array V2 analysis of the 7 samples. Total RNA from each sample was quantified using the NanoDrop ND-1000. The sample preparation and microarray hybridization were performed based on the Arraystar's standard protocols. Briefly, total RNAs were digested with Rnase R (Epicentre, Inc.) to remove linear RNAs and enrich circular RNAs. Then, the enriched circular RNAs were amplified and transcribed into fluorescent cRNA utilizing a random priming method (Arraystar Super RNA Labeling Kit; Arraystar). The labeled cRNAs were hybridized onto the Arraystar Human circRNA Array V2 (8x15K, Arraystar). After having washed the slides, the arrays were scanned by the Agilent Scanner G2505C. Agilent Feature Extraction software (version 11.0.1.1) was used to analyze acquired array images. Quantile normalization and subsequent data processing were performed using the R software limma package. Differentially expressed circRNAs with statistical significance between two groups were identified through Volcano Plot filtering. Differentially expressed circRNAs between two samples were identified through fold change filtering. Hierarchical clustering was performed to show the distinguishable circRNA expression pattern among samples.