High-Fidelity DNA Polymerase

The fidelity of a DNA polymerase is the result of accurate replication of a desired template. The fidelity of DNA polymerase has multiple steps; the ability to read a template strand, select the correct nucleoside triphosphate and insert the appropriate nucleotide at the 3’ primer terminus.

To effectively discriminate correct vs. incorrect nucleotide incorporation, some DNA polymerases possess a 3’ to 5’ exonuclease activity. This process, known as “proofreading,” is used to excise incorrectly incorporated mononucleotides, which are then replaced with the correct nucleotides. High-fidelity PCR uses DNA polymerases that couple low misincorporation rates with proofreading to give faithful replication of the target DNA of interest.

Fidelity is essential for processes in which the DNA sequence must be correct following amplification.

For example:
- Cloning/subcloning DNA for protein expression
- SNP analysis
- Next generation sequencing applications

Genie Fusion Ultra High-Fidelity DNA Polymerase is a next generation enzyme for robust PCR with ultra high-fidelity. With extremely low error rates x53-fold lower than Taq, Genie Fusion Ultra HighFidelity DNA Polymerase sets a new level of performance for standard and long fragment PCR amplification.

Employing a unique extension factor as well as specificity-promoting and plateau-inhibiting factors Genie Fusion greatly improves long range amplification ability, specificity, and PCR yield. Genie Fusion is capable of amplifying long fragments such as 40 kb λ DNA, 40 kb plasmid DNA, 20 kb genomic DNA and 10 kb cDNA. In addition, Genie Fusion has a excellent resistance to PCR inhibitors and can be used for direct PCR amplifications of bacteria, fungi, plant tissues, animal tissues, and even whole blood samples. Genie Fusion contains two monoclonal antibodies inhibiting the 5´→3´ polymerase activity and 3´→5´ exonuclease activity at room temperature which enables Genie Fusion to perform superspecific Hot-Start PCRs. The amplification generates blunt-ended products, which are compatible with GenieClone DNA Assembly Cloning Kit (Assay Genie, #MORV0102)

Figure: Using mouse tail lysate as a template, a target fragment of 2.5 kb was amplified with Genie Fusion Ultra High-Fidelity DNA Polymerase, a high-fidelity DNA polymerase from company A, and a high-fidelity DNA polymerase from company B. The Tm of all primers are approximately 60℃ (calculated in Primer Premier 5). ( M: 5kb DNA Marker, 1: Tomato leaf, 2: Rice leaf, 3: Polished rice, 4: Purified genomic DNA from rice leaf.)

• Ultra high-fidelity DNA polymerase with error rates x53-fold lower than conventional Taq and x6-fold lower than Pfu.
• Excels at amplifying long fragments such as 40 kb λ DNA, 40 kb plasmid DNA, 20 kb genomic DNA and 10 kb cDNA.
• Superior performance with GC-rich templates.
• Extreme resistance to PCR inhibitors.
• Excellent choice for direct PCR from bacteria, fungi, plant tissues, animal tissues and whole blood samples.
• Formulated with dual monoclonal antibodies for super specific Hotstart PCR.
• Compatible with GenieClone DNA Assembly Cloning Kit (MORV0102).