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. 2021 Apr 17;184:113218. doi: 10.1016/j.bios.2021.113218

Fig. 1.

Fig. 1

Overview of dWS-CRISPR assay. (a) One-pot WS-CRISPR reaction mixture is first prepared in one tube. After distributed into QuantStudio 3D digital chip, over ten thousand sub-nanoliter (~0.7 nl) microreactions are isolated in microwells. When incubated at 52 °C, each microreaction with SARS-CoV-2 RNA target undertakes WS-CRISPR reaction and generates strong green fluorescence (positive spots), whereas not in those without target (negative spots). Scale bar is 300 μm. Through detecting and counting the positive microreactions (or spots), SARS-CoV-2 RNA can be quantified based on the proportion of positive spots. (b) Working principle of one-pot WS-CRISPR assay for SARS-CoV-2 detection. The WS-CRISPR reaction mixture contains Cas12a-crRNA complex, six DAMP primers (two outer primers of FO and RO, two inner primers of FI and RI, and two competition primers of FC and RC), ssDNA-FQ reporter, SuperScript IV reverse transcriptase, Bst DNA polymerase, and SARS-CoV-2 RNA in a one-pot format.