circRNA prediction, sequence determination, and validation.

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circRNA prediction, sequence determination, and validation. circRNA prediction, sequence determination, and validation. (A) Strategy for the identification of circRNAs and determination of their sequence and expression in cortical cell types. (B) DNA gel electrophoresis upon PCR amplification of 10 circRNAs identified as in (A). Full gels are available in Fig S2. Note the detection of bands with a molecular weight consistent with the expected exonic, but not intronic, regions. (C) Graph representing the cycle threshold (Ct) values upon RT-PCR of total RNA from the E14.5 mouse cortex, with or without a prior treatment with RNase R (as indicated) and using divergent primers for 30 predicted circRNAs together with linear and circular controls (red dashed line). Dashed black line indicates the Ct threshold of detection. N = 2, n = 3; bars = SDs (*P < 0.05; **P < 0.01; ***P < 0.001; t test). (D, E) Distribution of CiCo’s entries overlapping circRNAs deposited in circBase and (E) Venn diagram representing the proportion of circBase- and CiCo-specific circRNAs as well as common ones. Martina Dori et al. LSA 2019;2:e201900354 © 2019 Dori et al.