Table 2.
Gene targets, primers and probes used in multiplex on the QuantStudio 7 * platform to detect B. anthracis and B. cereus biovar anthracis (Bcbva) using real-time PCR.
| Target | Oligo | Primer/Probe Sequence (5′–3′) | Final Concentration (nM) |
|---|---|---|---|
| Ba-1 | Forward | GTACATCTTCTAGCTGTTGCAA | 900 |
| Reverse | ACGTAGGAAGACCGTTGATTA | 900 | |
| Probe | VIC-CGTTGTTGTGTATTTG-MGB | 250 | |
| capB | Forward | TAAGCCTGCGTTCTTCGTAAATG | 600 |
| Reverse | GTTCCCAAATACGTAATGTTGATGAG | 600 | |
| Probe | NED-TTGCAGCGAATGAT-MGB | 300 | |
| lef | Forward | CACTATCAACACTGGAGCGATTCT | 600 |
| Reverse | AATTATGTCATCTTTCTTTGGCTCAA | 600 | |
| Probe | Cy5-AGCTGCAGATTCC-MGB | 250 | |
| GI4 | Forward | GGAGATATTAACAAGAGATGGATTGGA | 700 |
| Reverse | CAGTAGGCTTGTCTGCTCTAATAAAATT | 600 | |
| Probe | FAM-ACATGCCAGCGTTTTTTGCCTCTACACA-BHQ1 | 150 |
* QuantStudio 7 excitation filter wavelengths in nm: 470 ± 15, 520 ± 10, 549.5 ± 10, 580 ± 10, 640 ± 10; emission filter wavelengths: 520 ± 15, 558 ± 12, 586.5 ± 10, 623 ± 10, 682 ± 10, 662 ± 10.