( rely on chemical modification ...)HotMasterAn innovative Hot Start/Cold Stop technology for better ,,, PCR* results

rely on chemical modification or antibodies to inhibit Taq polymerase; thus, their action is limited to the first cycle of the PCR reaction. In addition, they impart some disadvantages such as long pre-incubations at high temperatures
or protein contamination.

Eppendorf has developed an innovative Hot Start technology. This new technology uses a ligand that binds to the Taq polymerase in a temperature-dependent manner and inhibits it. This ligand is thermostable, and it is active in all cycles of the PCR. The net result is that Eppendorf's HotMaster kit not only offers Hot Start activity during the first cycle of the PCR, but also Cold Stop activity at temperatures below 60C during the annealing step of each and every cycle of the PCR.

The buffer is formulated to adjust the Mg⁺⁺ concentration automatically so that there is never a need for optimization of this critical component. It does this by weakly chelating Mg⁺⁺ ions: when Mg⁺⁺ is present in excess, it is bound by the chelating agent, but as it is needed by the reaction (for Taq or DNA), it is released. This is an innovative technology that further improves the sensitivity and specificity of HotMaster.

The series of experiments in this paper were designed to show that Eppendorf Hot Start/Cold Stop and self-adjusting Mg⁺⁺ technology offer a superior alternative to other Hot Start Taq polymerases. It increases specificity as well as the length of the product that is amplified relative to other Hot Start polymerases. Eppendorf's HotMaster also gives increased sensitivity: we were able to reproducibly amplify a single-copy gene from as little as 10 pg of human genomic DNA (about 2 genomic equival
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