Table 1.
Primers and Probes for RT-PCR
| Gene | Oligonucleotide | Primer/probe | Vendor/Cat.No. | Sequence 5’ to 3’ | 20X Stock Conc. | Final 1x conc. in the reaction |
|---|---|---|---|---|---|---|
| Subgenomic Envelope (E) | sgLeadSARSCoV2-F | Lead Forward Primer | IDT / custom DNA oligo | 5’-CGATCTCTTGTAGATCTGTTCTC | 17 μM | 0.85 μM |
| E_Sabeco_R | Reverse Primer | IDT / 10006890 | 5’-ATATTGCAGCAGTACGCACACA | 17 μM | 0.85 μM | |
| E_Sarbeco_P1 | Probe | IDT / 10006892 | 5’-/FAM/ACACTAGCCATCCTTACTGCGCTTCG/3IABkFQ/ | 6 μM | 0.30 μM | |
| Subgenomic Nucleocapsid (N) | sgLeadSARSCoV2-F | Lead Forward Primer | IDT / custom DNA oligo | 5’-CGATCTCTTGTAGATCTGTTCTC | 10 μM | 0.5 μM |
| wtN_R | Reverse Primer | IDT / custom DNA oligo | 5’-GGTGAACCAAGACGCAGTAT | 10 μM | 0.5 μM | |
| wtN_P | Probe | IDT / custom Prime Time qPCR probe) | 5’-/SUN/TAACCAGAATGGAGAA/ZEN/CGCAGTGGG/3IABkFQ/ | 5 μM | 0.25 μM | |
| Mouse RNase P (Rpp30) | Mouse_Rpp30_F | Forward Primer | IDT / custom DNA oligo | 5’-TCCAGTGTGCAAGAAAGCTAAATG | 1 μM | 0.05 μM |
| Mouse_Rpp30_R | Reverse Primer | IDT / custom DNA oligo | 5’-GGCAGTGCGTGGAGACTCA | 1 μM | 0.05 μM | |
| Mouse_Rpp30_P | Probe | Thermo Fisher / custom TaqMan™ Probe | 5’-/JUN/AGGGCTGAGACGAGTC/QSY/ | 0.5 μM | 0.025 μM |