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PCR / qPCR Reaction Mix Calculator (Master Mix)
Enter the volume per tube, the number of reactions and your kit's master mix concentration (for example 2× SYBR), and get the total mix, the master mix volume and the volume left for primers, template and water, with 10% extra already added for pipetting loss.
Inputs
μL
×
Results
- Total mix (×1.1)
- 220 μL
- Master Mix
- 110 μL
- Primer/template/H₂O
- 110 μL
Formula
Total (μL) = Rxn vol × N × 1.1
MM (μL) = Rxn vol ÷ MM× × N × 1.1
Tip: Auto ×1.1 for 10% pipetting loss · MM ratio per kit (e.g. 2× SYBR)
You got this step right, but a whole experiment has dozens of linked parameters. Change one dilution factor and everything downstream has to be recalculated. In Vara the parameters of your whole experiment are connected, so changing one recalculates the rest automatically. Manage 3 experiments for free.
FAQ
Why prepare 10% extra?+
Each transfer loses a little liquid on the tip and tube wall, and viscous master mix makes this worse. Preparing reactions × 1.1 ensures the last tube is not short. For very small runs (2 or 3 reactions), add one extra reaction manually instead.
What does 2× master mix mean, and how much do I add?+
2× means the kit's premix is twice concentrated, so it takes up half of the final reaction volume. In a 20 μL reaction that is 10 μL of master mix per tube, with the other 10 μL made up of primers, template and water. A 5× premix takes up one fifth; the calculator scales by the factor you enter.
How do I split the remaining volume between primers, template and water?+
The calculator reports the volume left after the master mix. Divide it following your kit's instructions: typical qPCR final concentrations are 0.2 to 0.5 μM per primer, with 1 to 2 μL of DNA or cDNA template per tube, and nuclease-free water to volume. Keep the template amount the same across a batch.