Build a PCR master-mix volume plan from per-reaction component volumes. Add the number of samples, controls, technical replicates, and a pipetting overage to calculate total reactions to prepare, master-mix volume, template volume, and the amount of each reagent required.
Required reactions = samples × replicates + controls. Prepared master mix uses required reactions × (1 + overage). Water per reaction = reaction volume − template − primers − PCR mix. Template is calculated only for actual reactions because it is normally added separately.
Use the main result as a planning estimate and read the supporting metrics to understand which assumptions drive it.
Results depend on the entered assumptions. Confirm critical decisions with measured operational data or a validated laboratory protocol.
For 24 samples in duplicate plus 2 controls, there are 50 reactions. At 20 µL each with 2 µL template, 1 µL primers, 10 µL PCR mix, 7 µL water, and 10% overage, prepare 990 µL of master mix for 55 reaction-equivalents.
It uses only the values entered on the page and the formula shown above. Review the inputs whenever operating conditions or lab protocols change.
Use measured averages where possible and include the provided overage, utilization, overlap, or efficiency input rather than hiding variability in another field.
Zero is accepted only where it has a meaningful interpretation. The calculator blocks zero denominators and other combinations that cannot produce a valid result.
No. It is a planning aid. Confirm reagent compatibility, instrument limits, assay performance, and pipetting feasibility with your validated protocol.
Keep full precision through the calculation and round only the quantities you need to measure or report. The calculator follows that approach internally.
| Input | Unit | Role |
|---|---|---|
| Samples | samples | Distinct biological samples |
| Control reactions | reactions | Positive, negative, or no-template controls |
| Replicates per sample | × | Technical reactions per sample |
| Final reaction volume | µL | Total volume in each well |
| Template per reaction | µL | Template added separately |
| Combined primers per reaction | µL | Forward plus reverse primer volume |
| PCR master mix per reaction | µL | Commercial mix or enzyme/buffer blend |
| Pipetting overage | % | Extra master mix for transfer loss |