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The Arraystar human circRNA Array analysis

ZD Zhao-Ru Dong
AK Ai-Wu Ke
TL Tao Li
JC Jia-Bing Cai
YY Ya-fei Yang
WZ Wei Zhou
GS Guo-Ming Shi
JF Jia Fan
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Total RNA from each sample was quantified using a NanoDrop ND-1000. The sample preparation and microarray hybridization were performed based on Arraystar’s standard protocols. Briefly, total RNA from each sample was amplified and transcribed into fluorescent cRNA utilizing random primers according to Arraystar’s Super RNA Labeling protocol (Arraystar Inc.). The labelled cRNAs were hybridized onto the Arraystar Human circRNA Array. After washing the slides, the arrays were scanned by an Axon GenePix 4000B microarray scanner.

Scanned images were then imported into GenePix Pro 6.0 software (Axon) for grid alignment and data extraction. Quantile normalization and subsequent data processing were performed using the R software package. Differentially expressed circRNAs with statistical significance between two groups were identified through volcano plot and fold change filtering. Hierarchical clustering was performed to show the distinguishable circRNA expression pattern among samples.

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