Table 2.
Reagent | Steps | Well coverage | Total travel (min) | Total Reaction hold (min) | Total drain (min) | Total cycle (min)/step |
---|---|---|---|---|---|---|
TCA | 3 | 144 | 2.67 | 0.00 | 0.05 | 2.72 |
Activatora | 1 | 96 | 1.07 | 0.67 | 0.02 | 1.76 |
Cap | 1 | 96 | 1.07 | 0.33 | 0.02 | 1.42 |
Oxidizer | 1 | 96 | 1.07 | 0.33 | 0.02 | 1.42 |
Wash | 5 | 144 | 4.45 | 0.00 | 0.08 | 4.53 |
Total cycle time (min) | 11.85 |
Here it is shown how the total synthesis cycle time on the NG-1536-AMOS platform is calculated. For example, during the deblock step, TCA is delivered to three rows simultaneously covering 144 wells. The total time it takes the titer plate to move continuously over all 32 rows while delivering TCA (e.g. 3.5 μl/well) is 0.89 min. After a reaction hold period of 0.00 s, TCA is then drained from the wells (vacuum-to-waste valve is open for 1 s). If all these steps are repeated two more times, it will take 2.72 min to complete the deblock portion of the synthesis cycle. Coupling, capping and oxidation steps are also factored in the same way to derive at 11.85 min total cycle time.
This step includes phosphoramidite addition (activator + base per coupling step). The washing reagent “Wash” used for synthesis is acetonitrile.