Magbead viral dna rna kit

For detecting the viral load in the supernatant, SARS-CoV-2 RNA was isolated by Magbead Viral DNA/RNA Kit (CWBIO), and SARS-CoV-2 nucleic acid detection kit (Da an Gene Co., Ltd.) was used to quantify viral RNA. For detecting viral RNA or other gene expression in cells, the total RNA was extracted with TRIzol reagent (Thermo Fisher, USA) and reverse-transcribed into cDNA using PrimeScript RT Master Mix (Takara, RR036A) according to the manufacturer’s instructions. RT-qPCR was performed with PowerUp SYBR Green Master Mix (Thermo Fisher, A25742) on the ABI QuantStudio5 (Applied Biosystems, USA). The relative abundance of target RNA was normalized to the human housekeeping gene actin beta ACTB. The primer sequences for detecting SARS-CoV-2 genome or other genes were shown in Supplementary Material Table S1.

For SARS-CoV-2 RNA quantification, RNA was isolated by Magbead Viral DNA/RNA Kit (CWBIO). SARS-CoV-2 nucleic acid detection kit (Da’an Company) was used to detect the virus. The limit of detection (LOD) of this kit reached 0.5 copies/μL. For the detection of the viruses and cytokines in mouse tissues, total RNA was isolated from tissue samples with TRIzol per the manufacturer’s instructions. mRNA was reverse transcribed into cDNA by PrimeScript RT reagent Kit (Takara). The cDNA was amplified by a fast two-step amplification program using ChamQ Universal SYBR qPCR Master Mix (Vazyme Biotech Co., Ltd) or Taq Pro HS Universal Probe Master Mix (Vazyme Biotech Co., Ltd). Gapdh was used to normalize the input samples using the ΔCt method. The relative mRNA expression of each gene was normalized to Gapdh housekeeping gene expression in the untreated condition, and fold change was calculated by the ΔΔCT method relative to those in untreated samples. The qRT-PCR primers are listed in table S7.