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. 2022 Feb 21;10:845688. doi: 10.3389/fbioe.2022.845688

FIGURE 1.

FIGURE 1

The outline of CRISPR/Cas12a-E-LAMP detection system. In the CRISPR/Cas12a-E-LAMP method, the gDNA of S. flexneri was extracted and then used in the LAMP reaction (10 μl) at 60°C for 20 min and covered by 20-μl mineral oil. The Cas12a reaction reagents (20 μl) was pre-placed in the lid and mixed with LAMP products at 37°C for 20 min. Once the Cas12a find the PAM site and the sgRNA complementary pairing with the target DNA, Cas12a endonuclease is activated. Then, Cas12a endonuclease cis-cleavage the LAMP products and split the ssDNA-FQ reporter indiscriminately by trans-cleavage, generating the fluorescence signal visible to the naked eye under LED blue light.