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The TripleMaster PCR System for three different amplifications

urer specifications. The PCR volume was 50 l.

Result Experiment 2

The TripleMaster PCR System shows a degree of accuracy that is normally only achieved using products that are specifically designed for high-fidelity applications. Although the tested PfuTurbo System (Stratagene) is revealed to have the lowest error rate, it is suitable only for the amplification of very short sequences (< 200 bp) see Fig. 2. Table 2: Final concentrations The new Eppendorf High Fidelity Buffer (short range) was used in the preparation of the TripleMaster PCR System. Fig. 2: Direct comparison of the fidelity of DNA polymerases under real PCR conditions using relative data Experiment 3: TripleMaster PCR System in a test with extremely GC-rich templates

To test the performance of TripleMaster PCR System during the amplification of complex templates, a GC-rich (> 70 %) 490 bp fragment from the human -actin gene was amplified.

The intron sequence between exon 1 and exon 2 of the human -actin gene has 85 % GC content and contains stable secondary structure elements. Table 3: Final concentrations in 50 l reaction Result Experiment 3

Taq DNA polymerase with 2 % DMSO is unable to amplify such a complex template.

However, the TripleMaster PCR System produces very good results either in combination with only 2 % DMSO or with only Eppendorf TaqMaster (1x) see Fig. 3.
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