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    Cat. No.
    Size
    List Price*
    QTY
    BIO-21065
    500 Units
    $380.00
    +
    *To check your pricing please Login or contact us

    Description

    BIO-X-ACT™ Short DNA Polymerase is a high-performance enzyme designed for difficult or problematic PCR applications.

    BIO-X-ACT Short is recommended for short genomic DNA fragments of up to 3 kb. When using Lambda DNA as template, the best performance is achieved within the 100 bp-5 kb range.

    Product Highlights

    • High performance - for templates that fail with standard Taq DNA Polymerases
    • For problematic templates - can work with high GC content, dirty templates or difficult melting profiles
    • Good for short amplicons - amplifies genomic fragments up to 3 kb
    • Flexible format - Available as a ready-to-use 2x reaction mix (BIO-X-ACT™ Short Mix)

    Product Description

    BIO-X-ACT™ Short DNA Polymerase is a high-performance enzyme designed for difficult or problematic PCR applications. BIO-X-ACT is the polymerases of choice for difficult applications that would normally fail with standard Taq polymerases (fig. 1).

    BIO-X-ACT Short is recommended for short genomic DNA fragments up to 3 kb and Lambda DNA up to 5 kb.

    For enhanced specificity, BIO-X-ACT Short is supplied with a vial of Hi-Spec Additive. Hi-Spec Additive is a very efficient enhancer, which helps to reduce the formation of false background bands and smearing. The specificity and performance can be further improved with the use of 3% DMSO (not supplied), which is designed for GC or AT-rich DNA, "dirty" templates or sequences with difficult melting profiles.

    Applications

    • Suitable for TA cloning
    • GC-rich templates
    • Crude sample PCR
    • Forensic applications
    Main

    PCR Selection Chart

    Select the best reagent for your research

    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

    Specification

    Components

     Reagent

    500 Units

    BIO-X-ACT Short DNA Polymerase

    125 µL

    10x OptiBuffer

    2 x 1.2 mL

    50 mM MgCl2 Solution

    1.2 mL

    5x Hi-Spec Additive

    1.2 mL

    Concentration

    4u/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.

    Shipping conditions

    On Dry Ice or Blue Ice.

    One unit will incorporate 10 nmoles of dNTPs in 30 min at 72°C.


    BIO-X-ACT is a trademark of Bioline.



    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.

    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.