Table 1.
Components for one reaction with a single nucleotide triphosphate concentration.
| Initial Concentration | Final concentration* | Volume (μL) | |
|---|---|---|---|
| Primer/Template | 500 nM | 100 nM | 7 |
| DNA polymerase | 5 nM | 1 nM | 7 |
| 5× assay buffer | 5X | 1X | 7 |
| Water | 7 | ||
| Total | 28 | ||
| Take out for zero point | −4 | ||
| Total | 24 | ||
| Initiate with 2AP-TP or 4CIN-TP | 6 | ||
| 30 | |||
*
The final concentration column in Table 1 is calculated assuming the nucleotide has been added as in Step 10, i.e. the final 30 μL volume solution has been mixed and the assay has started.