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. 2022 Mar 1;2:834655. doi: 10.3389/fbinf.2022.834655

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Copyright © 2022 Vromman, Anckaert, Vandesompele and Volders.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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CIRCprimerXL primers are efficient and can validate circRNAs. (A) The efficiency (E) of the primers is assessed using a 6-point 10-fold dilution of a synthetic DNA template positive control. All primers showed an E-value between 1.9 and 2.1, which corresponds to efficiency between 90 and 110%. (B) These primers where then tested on SW480 total RNA (two treatment replicates, two qPCR replicates). The 20 circRNAs remained stable (similar Cq value) upon RNase R treatment, in contrast to the 4 linear control genes, that were degraded (increase in Cq value).