Table 3.
Final optimized PCR chemistry for various genes in a final volume of 150 µL. All reagents were obtained from BioFire Defense, Inc.TM for quality of final optimization prior to lyophilization.
| Reagent | Starting Concentration | E. coli | Salmonella | Aeromonas | 16S |
|---|---|---|---|---|---|
| Stabilization buffer | 4X | 1X | 1X | 1X | 1X |
| MgCl2 with BSA | Variable | 3.0 mM | |||
| 2.5 mM each dNTP’s | 25 mM | 200 nM | 200 nM | 200 nM | 200 nM |
| TaqStart Antibody | 5 µg/µL | 0.4 | 0.4 | 0.4 | 0.4 |
| Forward Primer | 100 µM | 900 nM | 100 nM | 1000 nM | 500 nM |
| Reverse Primer | 100 µM | 900 nM | 100 nM | 1000 nM | 500 nM |
| TaqMan Probe | 100 µM | 200 nM | 100 nM | 250 nM | 100 nM |
| Vtaq DNA polymerase Glycerol Free | 5 U/µL | 1.8 µL | 1.8 µL | 1.8 µL | 1.8 µL |
| Water | Variable | Variable | Variable | Variable | Variable |