Nanodrop 1000 spectrophotom

The DNA targets of WT S gene and MT SARS-CoV-2 (covering D614G, N501Y and 69/70 deletion sites) were synthesized (Table S4) and then cloned into pUC57 plasmid.
RNA targets were prepared through in vitro transcription with a HiScribe T7 High-Yield RNA Synthesis Kit. Briefly, RNA transcription templates containing a T7 promoter were amplified from pUC57-WT-DNA and pUC57-MT-DNA with specific primers (Table S2) using the NEBNext Q5 Hot Start HiFi PCR Master Mix according to manufacturer's instruction, followed by purification using a Gel Extraction Kit. In vitro transcription was performed in 20 μL reaction volume, according to a standard RNA synthesis protocol, at 37 °C for 16 h. Finally, RNA transcripts were treated with DNaseI to remove transcription templates, purified with an RNA Cleanup Kit, and quantified using Thermo Fisher Nanodrop 1000 Spectrophotom.

LbCas12a, HiScribe T7 High-Yield RNA Synthesis Kit, RNA Cleanup Kit, NEBNext Q5 Hot Start HiFi PCR Master Mix, NEBuffer2.1 were ordered from NEB (Beijing, China). DNaseI, recombination RNase inhibitor (RRI), PrimeSTAR max were ordered from Takara Bio (Beijing, China). T-vectors, Pfu DNA polymerase were ordered from TIANGEN Biotechnology (Beijing, China). One-Step RT-PCR Kit was ordered from Vazyme Biotech (Nanjing, China). Gel Extraction Kit was ordered from Omega Biotech (Shanghai, China). Lateral flow strips were purchased from Magigen Biotech (Guangzhou, China). Oligonucleotides were synthesized by GENEWIZ (Jiangsu, China). S gene DNA targets were synthesized by Generay Biotech (Shanghai, China). Nucleic acid was quantified using Thermo Fisher Nanodrop 1000 Spectrophotom. Fluorescence signals were recorded with Tecan's Spark 20M.