PCR

PCR master mix

A component table for any number of tubes, with presets for PCR, RT and colony PCR

Reaction preset Number of reactions Volume per reaction µl Overage for losses % Template is added separately

Per line: the component name, its stock concentration, the final concentration and a unit. For a component added at a fixed volume, such as an enzyme, leave the stock column empty and put the volume in the third column with the unit ul.

—Total master mix

—Master mix per tube

—Template per tube

—Reactions + overage

Each volume follows from C₁V₁ = C₂V₂. Template is deliberately kept out of the mix because it differs per tube; untick that option if every tube takes the same template. Set the overage to at least ten per cent, and more beyond twenty tubes — loss on the pipette tip adds up faster than it looks.