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. 2022 Sep 10;5:947. doi: 10.1038/s42003-022-03894-9

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a Endonuclease activity of Cas12a. Lane Marker: DL1000 (Takara Biomedical Technology (Beijing) Co., Ltd., China); Lane a1: Cas12a + Cs_crRNA + ITS2 fragments of Crocus sativus; Lane a2: Cas12a + Cs_crRNA: Lane a3: ITS2 fragments of Cr. Sativus. b Collateral cleavage activity of Cas12a. Lane b1: Cas12a + Cs_crRNA + ITS2 fragments of Cr. sativus + 50 bp ssDNA; Lane b2: 50 bp ssDNA; b3: Cas12a + Cs_crRNA + 50 bp ssDNA. c Fluorescence signal of GAGE. The two groups were incubated at 37 °C for 25 min and detected by excitation with 470 nm light. Cs: Cas12a + Cs_crRNA + ITS2 fragments of Cr. sativus + ssDNA reporter; CK (negative control): Cas12a + Cs_crRNA + H₂O + ssDNA reporter. d Specificity of GAGE. The plant materials in the circle are the stigma of saffron. The DNA substrate of two groups were as follows: Cs: ITS2 fragments of Cr. sativus and CK (negative control): nuclease-free water. e Sensitivity of GAGE. The seven groups had different concentration of DNA substrates (ITS2 fragments of Cr. sativus), e1: 10 ng/μL; e2: 1 ng/μL; e3: 0.1 ng/μL; e4: 0.01 ng/μL; e5: 0.001 ng/μL; e6: 0.0001 ng/μL and CK (negative control): 0 ng/μL. All plots data represented means +/− standard deviation (SD) from three independent replicates.