PCR & qPCR calculators
The arithmetic of amplification: setting up a reaction and its master mix, choosing an annealing temperature and planning a gradient, sizing the product, and diluting a template to the amount you want per reaction.
For quantitative PCR there are tools for summarising replicate Ct values, expressing a target relative to a reference gene by the ΔCt, ΔΔCt and efficiency-corrected (Pfaffl) methods, measuring amplification efficiency from a dilution series, and quantifying unknowns against a standard curve. Each states its assumptions, because a qPCR number is only as good as the assay behind it.
PCR Master Mix CalculatorComponent volumes for any number of reactionsOpen calculatorqPCR ΔΔCt CalculatorRelative gene expression by the 2^−ΔΔCt methodOpen calculatorPCR Product Size CalculatorAmplicon length from primer positions, with 5′ tailsOpen calculatorPCR Annealing Temperature CalculatorOptimal annealing temperature from primer Tm and productOpen calculatorGradient PCR PlannerTemperature of each column in a gradient blockOpen calculatorqPCR Replicate Ct CalculatorMean, SD and fold spread of replicate Ct values, with flagsOpen calculatorqPCR ΔCt CalculatorΔCt and relative expression to a reference geneOpen calculatorqPCR Efficiency-Corrected Ratio (Pfaffl)Fold change when target and reference efficiencies differOpen calculatorqPCR Efficiency CalculatorAmplification efficiency from a dilution series or a slopeOpen calculatorqPCR Absolute Quantification CalculatorCopy number of unknowns from a standard curveOpen calculatorPCR Template Dilution CalculatorDilute a template to the amount you want per reactionOpen calculator
Related equipment
Service documentation, failure modes and parts for the instruments these calculations are used with.