Accura® High-Fidelity Polymera

Accura® High-Fidelity Polymerase is the perfect solution for your everyday amplification and cloning needs. Amplify your genes for cloning and protein expression applications with confidence while getting the most out of your laboratory budget.

The Accura High-Fidelity polymerase is a fusion protein, consisting of a DNA binding domain and a unique, thermostable polymerase derived from a bacteriophage. When used according to recommendations, Accura High-Fidelity polymerase results in robust amplification of normal, high GC% or otherwise difficult templates

Fidelity

Accura High-Fidelity Polymerase delivers the accuracy you need for cloning and protein expresssion, generating equivalent numbers of functional clones when compared to Phusion, and significantly more than when using Taq (Fig. 1).

Figure 1. A 700bp blue colored protein gene was amplified using manufacturer's recommended conditions for Phusion High-Fidelity DNA polymerase (Thermo-Fisher), Accura High-Fidelity Polymerase, and standard fidelity EconoTaq® (Lucigen). The amplicons were then cloned using the Expresso® Rhamnose Cloning and Expression system (Lucigen). Blue colonies represent functionally accurate amplification of the gene of interest, while white colonies represent non-functional genes with sequence errors due to low-fidelity amplification. Some white colonies may also be due to empty vector.Accura and Phusion show a similar percentage of correct genes, both significantly better than compared to Taq polymerase.

Flexibility

Accura™ High-Fidelity Polymerase kits contain everything you need to obtain your amplicon of interest. HF buffer ensures the highest yield and fidelity, but for high GC % or difficult to amplify targets, use the included GC buffer to achieve results where other polymerases fail or produce spurious bands (Fig. 2).

Figure 2. Amplification of Human DNA “Tough Template” targets with elevated GC content of 58-78%. Sequences were amplified using published primers (Zhang (2009) Biotechniques 47:775-8) from purified human genomic DNA. Accura High-Fidelity Polymerase and Phusion were compared using their respective GC reaction buffers and manufacturer recommended conditions.

Performance

Easily amplify any standard gene for your day-to-day cloning and protein expression with Accura High-Fidelity DNA Polymerase using normal cycling conditions. When used according to recommendations, Accura High-Fidelity polymerase results in robust amplification of normal, high GC% or otherwise difficult templates. Accura routinely amplifies targets of up to 5 kb; larger targets of up to 10 kb have been achieved with optimization.

Figure 3. A) Amplification of two separate 3 Kb targets (polA and lacZ) using Accura High-Fidelity Polymerase with HF buffer and standard thermal cycling conditions. B) Amplification of a 5 Kb human DNA target (BRCA1).

Accura and Expresso: The fastest path to protein expression

Combine the fidelity of Accura High-Fidelity DNA polymerase with the speed of Lucigen's Expresso Cloning and Expression kits to amplify and clone your gene in less than 2 hours from start to finish, with recombinant clones by the next day. For short (up to 1 Kb), easy to amplify genes, cycling conditions can be shortened to approximately 30 minutes with Accura High-Fidelity polymerase. No PCR cleanup is required prior to cloning with Lucigen's no-enzyme, recombinational cloning system. Expresso takes just a few seconds to create perfect clones without sequence scars or extra unwanted amino acids. Expressing your protein of interest has never been simpler (Fig. 4).

Figure 4. A 700bp blue colored protein gene was amplified using Accura™ High-Fidelity Polymerase and HF buffer using an abbreviated PCR protocol (94°C initial denaturation for 1 minute, 25 cycles of 94°C for 5 seconds, 60 °C for 5 seconds, and 72 °C for 10 seconds, followed by a 1 minute final extension at 72°C). The amplicons were then cloned using the Expresso® Rhamnose Cloning and Expression system (Lucigen). Blue colonies represent functionally accurate amplification of the gene of interest, while white colonies represent non-functional genes with sequence errors due to low-fidelity amplification. Some white colonies may also be due to empty vector. Fidelity is decreased slightly, but a high percentage of all clones are still functional despite a <30 minute cycling time.