A simple magnetic nanoparticles-based viral RNA extraction method for efficient detection of SARS-CoV-2

Zhen Zhao; Haodong Cui; Wenxing Song; Xiaoling Ru; Wenhua Zhou; Xuefeng Yu

The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.02.22.961268 doi: bioRxiv preprint reaction solution was set up containing 15 μL of eluted supernatant, 7.5 μL of 4 × reaction buffer, 1.5 μL enzyme mixture, and optimized concentrations of primers (from 0.25 μM to 1 μM)and probe (from 0.25 μM to 1 μM). Subsequent thermal cycling was performed at 55°C for 15 min for reverse transcription, followed by 95°C for 30 s and then 45 cycles of 95°C for 10 s, 60°C for 35 s. Fluorescence readout were taken after each cycle, and the threshold cycle (Ct) was calculated by the Bio-Red analysis system based on plotting against the log10 fluorescence intensity. The oligonucleotide primers and probes were synthesized by GENEWIZ (Suzhou, China), and corresponding sequences are shown in Table 1 . Table 1 . Sequences of primers and probes for RT-PCR.