Optimization of the rapid sample lysis protocol.

(a) Representative plot of fluorescence intensity versus time for lysis buffer screening using pseudovirus containing fragments of N gene of SARS-CoV-2 (left panel). Fluorescent signal was obtained at 10 minutes for the Cas12a reaction (right panel). Error bars represent the mean ± SD, where n = 3 replicates. The pseudovirus was lysed at 80°C for 5 minutes. Conditions: 1, 100 mM TCEP + 0.5% Triton X-100; 2, 0.5% Triton X-100 + 0.5% Tween-20; 3, 0.5% Triton X-100 + 0.5% Tween-20 + 1 mM EDTA; 4, 800 mM guanidine hydrochloride + 0.5% Triton X-100 + 1 mM EDTA; 5, 800 mM guanidine hydrochloride + 0.5% Triton X-100 + 0.5% Tween-20. (b) Real-time (left panel) and end point (right panel) fluorescence detection for N gene of SARS-CoV-2. Viruses were lysed at 80°C (5-heated) or room temperature (5-RT) for 5 minutes in a buffer containing 800 mM guanidine hydrochloride, 0.5% Triton X-100, and 0.5% Tween-20. Error bars represent the mean ± SD, where n = 3 independent repeats each with 3 technical replicates. Numerical source data underlying this figure can be found in S1 Data. Cas, CRISPR associated proteins; SARS-CoV-2, Severe Acute Respiratory Syndrome Coronavirus 2; SD, standard deviation.

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