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TripleMaster PCR SystemThe Ideal Choice for Long-Range PCR

for scientists seeking to overcome the limitations of standard PCR systems.1, 2, 3, 4

The TripleMaster PCR Systems Tuning Buffer has a unique zwitterionic formulation (patent pending) that adequately maintains pH at high temperatures (72 0C 94 0C), reducing template and amplicon degradation which is caused by depurination to a minimum. In addition, the TripleMaster Enzyme Mix combines the efficiency of Taq DNA Polymerase with the 3 5 exonuclease activity of a proofreading thermostable enzyme, along with a polymerase-enhancing factor that provides an extremely high extension rate and maximal proofreading assisted fidelity.5

Materials and Methods

Eppendorf TripleMaster PCR System
Invitrogen ThermalAceTM DNA Polymerase Kit
Stratagene EXLTM DNA Polymerase Kit
Roche Expand Long Template PCR System
Roche Expand 20 kbPlus PCR System
Genomic DNA isolated using the Eppendorf Perfect gDNA Blood Mini Kit
Eppendorf Mastercycler Gradient Thermal Cycler
FIGE Mapper Electrophoresis System (Bio-Rad)
OmniPur Agarose (EM Science)
0.5x TBE (Eppendorf)
Ethidium Bromide (Sigma)

The performance of the TripleMaster PCR System for long-range PCR was tested against other PCR systems specifically designed for long-range PCR. A 29 kb fragment of the human Factor VIII gene and 24 kb and 18 kb fragments of the human tPA gene were amplified in parallel reactions. All reactions were treated and set up in accordance with the manufacturers recommended specifications. For the Roche systems, the Expand 20 kbPlus PCR System was used to amplify the 29 kb and 24 kb targets and the Expand Long Template PCR System
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5. TripleMaster and Perfect gDNA Blood Mini Kit Team Up to Amplify Long gDNA Templates
6. Using the TripleMaster PCR System for Robust Amplification of GC-Rich DNA Templates
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11. Microdetermination of Phosphorus Using the SpectraMax Plus Microplate Spectrophotometer: Choice of Microplate, Cuvette, or Test Tube Assay Formats (MaxLine Application Note #24)
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