- Efficient – specifically developed to give reliable amplification of fragments between 10 kb and 25 kb in length
- Specific – incorporates an antibody-mediated hot-start enzyme that remains completely inactive during PCR set-up to prevent non-specific amplification
- Fast – rapid enzyme activation and highly efficient amplification support reduced time to results
- Flexible – ideal for amplifying a broad range of large fragments from even complex targets including human genomic DNA
- Accurate – possesses 3’ – 5’ proofreading exonuclease activity for increased fidelity, thereby enabling cloning of long PCR products
RANGER DNA Polymerase is recommended for all Long PCR applications RANGER is an easy-to-use, high performance enzyme, specifically developed to amplify fragments up to 25 kb in length. RANGER contains a unique combination of a high-performance DNA polymerase and novel buffer system that deliver the improved efficiency necessary for reliable amplification of longer amplicons.
RANGER is an antibody-mediated hot-start enzyme that eliminates non-specific amplification during reaction set-up. The inactivated enzyme does not possess polymerase activity, thereby preventing the non-specific amplification, such as primer-dimer formation, that often hinder Long PCR reactions from the start.
RANGER Mix includes specially formulated buffering system containing dNTPs, MgCl2 and enhancers at optimal concentrations, designed to minimize the requirements for PCR optimization, in turn reducing time to results and eliminating the cost of unnecessary repeats. Furthermore, RANGER dramatically reduces the time needed to set up reactions, thereby minimizing the risk of contamination. Greater reproducibility is ensured by the reduction in the number of pipetting steps that can lead to pipetting errors.
- Long PCR
- TA cloning
- Higher fidelity PCR amplification
Having struggled with PFU and other polymerases, I was extremely happy to try RANGER DNA polymerase and it worked for both 8.5 kb and 6.5 kb.
NWCR Institute, Bangor University, UK
Product SelectionPlease refer to the PCR Selection Chart to confirm the recommended product for your PCR application.
Certification of Analysis (COAs)
RANGER Mix, 2x
2x 1.25 mL
10 x 1.25 mL
- BIO-25051: 100 x 50 µL Reactions: 2 x 1.25 mL
- BIO-25052: 500 x 50 µL Reactions: 10 x 1.25 mL
Storage & Stability
All components should be stored at -20°C upon receipt for optimum stability. Repeated freeze/thaw cycles should be avoided. When stored under the recommended conditions and handled correctly, full activity of the reagents is retained until the expiry date indicated on the outer box label.
Components may also be stored at +4°C if required, although storage at -20°C is recommended. When stored at +4°C, reagents will remain stable for a period of 2 weeks from date of receipt.
Shipped on Dry Ice or Blue Ice
Bioline's mixes contain magnesium chloride at the following concentrations (please see table below). An additional tube of 50 mM MgCl2 solution is supplied with each mix to facilitate further optimization if required.
|Bioline Mix||Final Magnesium Concentration|
|ACCUZYME Mix||2.0 mM.|
|BioMix / BioMix Red||2.5 mM.|
|ImmoMix / ImmoMix Red||3.0 mM.|
|BIO-X-ACT Short Mix||2.0 mM.|
Please Note: We do not recommend the storage of our polymerases at -80 °C as ice crystals could form on the active site, which may affect or destroy the activity of the enzyme.
|No or low PCR yield||Enzyme concentration too low – increase the amount of enzyme in 0.5 U increments.|
|Primers degraded – check quality and age of the primers.|
|Magnesium concentration too low – increase concentration in 0.25 mM increments with a starting concentration of 1.75 mM.|
|Primer concentration not optimized. Titrate primer concentration (0.3-1 µM); ensuring that both primers have the same concentration.|
|Template concentration too low – Increase concentration of template.|
|Perform a positive control to ensure that the enzyme, dNTPs and buffers are not degraded and/or contaminated.|
|Multiple Bands||Primer annealing temperature too low. Increase annealing temperature. Primer annealing should be at least 5°C below the calculated Tm of primers.|
|Prepare master mixes on ice or use a heat-activated polymerase.|
|For problems with low specificity. Try adding 3% DMSO (not supplied) to improve specificity.|
|Smearing or artifacts||Template concentration too high. Prepare serial dilutions of template.|
|Too many cycles. Reduce the cycle number by 3-5 to remove non-specific bands.|
|Enzyme concentration too high - decrease the amount of enzyme in 0.5 U increments.|
|Extension time too long. Reduce extension time in 0.5-1 minute increments.|