Scale your PCR reactions with automatic overage. Customize components for any polymerase system.
| Component | Stock | Final | Per Rxn (µL) | Master Mix (µL) |
|---|---|---|---|---|
| Total Master Mix Volume | ||||
| + Template (added per tube) | — | |||
Set the number of reactions, volume per reaction, and overage percentage (10% is standard to account for pipetting losses). Click a polymerase preset to auto-fill typical component concentrations, or customize each row in the table. Template DNA is listed separately since it varies per tube and isn't part of the master mix.
The "Master Mix" column shows total volumes including overage — this is what you actually pipette when preparing the mix.
Always make a master mix when running multiple reactions. It reduces pipetting errors and ensures consistency across samples. Add the template last, after aliquoting master mix into individual tubes.
Overage: 10% extra is standard for up to ~20 reactions. For larger numbers of reactions (50+), you can reduce to 5% since pipetting errors average out.
Common issues: If your PCR fails, check MgCl₂ concentration first (too much causes nonspecific amplification, too little reduces yield). DMSO or betaine (3–5% final) can help with GC-rich templates. For colony PCR, reduce template volume and increase initial denaturation time to 10 minutes.