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    Cat. No.
    Size
    List Price*
    QTY
    BIO-21121
    250 Units
    $370.00
    +
    BIO-21122
    500 Units
    $676.00
    +
    *To check your pricing please Login or contact us

    Description

    Ideal for all Long PCR applications, including sequencing, mapping of chromosomal translocation breakpoints and other structural variations.

    Product Highlights

    • 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 blend 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

    Product Description

    RANGER DNA Polymerase is recommended for all long PCR applications. RANGER is an easy-to-use, high performance enzyme blend, specifically developed to amplify fragments up to 25 kb in length. RANGER contains a unique combination of a high-performance DNA polymerases and novel buffer system that deliver the improved efficiency necessary for reliable amplification of longer amplicons.

    RANGER is an antibody-mediated hot-start enzyme blend that eliminates non-specific amplification during reaction set-up. The inactivated enzymes do 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 is provided with a specially formulated reaction buffer containing dNTPs, MgCl2 and enhancers at optimal concentrations, minimizing the requirements for PCR optimization, in turn reducing time to results and eliminating the cost of unnecessary repeats.

    Applications

    • Long PCR
    • TA cloning
    • Higher fidelity PCR amplification
    Main
    Improved yield and specificity in Long PCR.

    PCR Enzyme Guide

    Download the PCR Enzyme Guide with detailed product descriptions and performance data to help you choose the best product for your research

    PCR Selection Chart

    Select the best reagent for your research

    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 Selection

    Please refer to the PCR Selection Chart to confirm the recommended product for your PCR application.


    Specification

    Components

    Reagent

    250 Units

    500 Units

    RANGER DNA Polymerase

    1 x 62.5 µL

    1 x 125 µL

    5x RANGER Reaction Buffer

    1 x 1.2 mL

    1 x 1.2 mL

    Volume

    • BIO-21121: 62 x 50 µL Reactions: 1 x 62.50 µL
    • BIO-21122: 125 x 50 µL Reactions: 1 x 125 µL

    Concentration

    • 4 u/µL

    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.

    RANGER DNA Polymerase may also be stored at +4°C for up to 2 weeks, although storage at -20°C is recommended.

    Shipping conditions

    On Dry Ice or Blue Ice.



    FAQs

    At Bioline we pride ourselves in supplying high-quality polymerases to suit your requirements. To aid your selection of the most suited enzyme for your specific applications, please see our enzyme selection tool.

    All our polymerases are guaranteed for a period of 12 months from the date of purchase. These should be stored at -20°C during this time for optimal retention of activity.

    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.

    PCR can be a challenging technique, with various parameters to optimize to achieve the best results. If you are having problems, these could be easily resolved by addressing a few issues. Please see our PCR troubleshooting guide for suggestions and help with your specific problems.

    Observation Recommended Solution(s)
    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.


    These terms refer to parameters to be considered when performing PCR and are important features in selecting the correct enzyme for your needs. Understanding what they mean is therefore crucial: Yield: The amount of DNA produced in a PCR reaction. Fidelity: The accuracy of the enzyme at incorporating the correct dNTP to the elongating DNA strand. Processivity: The length of time a polymerase is associated with the template and therefore the size of fragment which can be amplified. Specificity: A measure of the unwanted by-products generated in a reaction.

    During PCR setup at room temperature, standard polymerases will have some activity. When using a hot-start polymerase, this enzyme will have no activity at room temperature, thus reducing the risk of unspecific products in the reaction due to these mis-primed oligonucleotides. This feature makes this type of polymerase ideally suited to high-throughput applications, where the reactions may be left sitting at room temperature for prolonged periods of time.

    In nature, the ability of a DNA polymerase to correct misincorporations of nucleotides in the DNA strand being elongated is often crucial to the survival of the host organism. This ability is termed "proofreading activity" and occurs in the 3' to 5' direction. This activity also leads to the polymerase removing unpaired nucleotides overhanging at the 3'end (A-overhangs), creating blunt ends.

    One unit is defined as the amount of enzyme that incorporates 10 nmoles of dNTPs into acid-insoluble form in 30 minutes at 72°C.

    All Bioline polymerases are available in the convenient format of a 2x master mix, containing all the reagents and additives necessary to perform successful PCR, with the exception of template, primers and water. This formulation not only provides a convenient and hassle-free PCR setup, but also significantly reduces the chances of human error, inaccuracies and contamination by reducing the pipetting steps required.

    If possible, we would recommend an additional cleanup of affected samples, for instance, customers had very good experiences with the clean-up of samples containing humic acids with SureClean Plus. Alternatively decreasing the template concentration will help to minimize the amount of inhibitors. The amount of primer can be increased, but do not exceed the suggested limits.

    Please click here in order to request your sample. You will receive an email confirmation within two business days with delivery details.



    Reviews

    ""Having struggled with PFU and other polymerases, I was extremely happy ... "