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. 2021 Apr 27;12:649010. doi: 10.3389/fmicb.2021.649010

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Copyright © 2021 Huang, Gu, Xue, Yu, Tang, Huang, Zhang and Jiao.

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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Feasibility assessment of the CRISPR-Cas12b-based C. jejuni detection system. (A) The cis-cleavage activity of Cas12b. Lane M, DL1000 Marker. Lane 1: cleavage by Cas12b of the PCR product. In this system, the PCR product (substrate of Cas12b), sgRNA, Cas12b, and other reagents were added. Lane 2: negative control. Cas12b was not added in the system. (B) The trans-cleavage activity of Cas12b. Upper panel, Tube 1: all required components were added; Tube 2: the 5′−⁶FAM-N12-3′-BHQ1 probe was not included; Tube 3: only Cas12b was added; Tube 4: only template DNA was added; Tube 5: only the 5′−⁶FAM-N12-3′-BHQ1 probe was added. Lower panel: fluorescence intensity corresponding to each tube in panel (B) upper panel. Paired two-tailed t test, ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001. ND, Not detected. NC, Negative control.