Christian Rohrer, Eppendorf AG Introduction

Standard PCR components (Taq polymerase and reaction buffers) are used for the general amplification of relatively short DNA fragments (100 bp to roughly 2,000 bp).

However, demanding applications (such as long range PCR [1-2], high-fidelity PCR, or the amplification of complex templates) require additional reagents specifically designed for that application. In the past, separate kits would have been required for different applications. The new Eppendorf TripleMaster PCR System, however, is a universal solution for all of these applications.

Due to its unique composition of three different enzymes (Taq DNA polymerase, polymerase enhancing factor, and proofreading enzyme) and two innovative buffers (Tuning Buffer for long range PCR and High Fidelity Buffer for short range PCR), the TripleMaster PCR System produces excellent results for applications involving long range PCR, high-fidelity PCR, or amplification of complex templates.

Methods and results

Experiment 1: Amplification of very long fragments from DNA and genomic human DNA

In order to determine the performance of the TripleMaster PCR System, a 40 kb fragment from DNA and both a 27 kb and a 24 kb fragment from humangenomic DNA were amplified in two parallel preparations using the TripleMaster PCR System as well as leading PCR systems that are specially designed for long range PCR.

All PCR reactions were prepared in accordance with manufacturer protocols in 50 l volumes. Amplification was carried out in the Eppendorf M
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