Amplification of mitochondrial DNA using the REPLI-g Mitochondrial DNA Kit involves two basic steps. First, total DNA sample is denatured at 75°C in REPLI-g mt Reaction Buffer, then cooled. Next, REPLI-g DNA Polymerase is added for the isothermal amplification reaction.
0.1 ng total DNA (representing approximately 15 cells) containing approximately 100 fg of mitochondrial DNA was amplified using the REPLI-g Mitochondrial DNA Kit. The total yield of mitochondrial DNA after amplification was up to 4 µg, corresponding to a 4 x 107-fold increase in mitochondrial DNA.