BIOTAQ™ Red DNA Polymerase - Discontinued


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BIOTAQ™Red DNA Polymerase is a formulation of the regular BIOTAQ DNA polymerase, which contains a non-toxic and non-hazardous red dye.

The red dye provides easy and quick identification of reactions to which the enzyme has been added and facilitates visual confirmation of complete mixing.

Features & Benefits

  • Red color - easy visual recognition
  • Ready to use format - direct loading onto agarose gels
  • Good standard Taq polymerase - ideal for setting up new procedures
  • Easy to use - designed for easy optimization of PCR applications
  • High quality reagents - contains ultra-pure dNTPs manufactured by Bioline
  • Flexible format - supplied with 2x PolyMate Additive for difficult or "dirty" templates
  • Suitable for TA cloning - leaves 'A' overhang
  • Alternative format - available as a ready-to-use 2x reaction mix (BioMix™ Red)

Applications

  • Routine PCR applications
  • TA cloning
  • High throughput

Description

BIOTAQ™ Red DNA Polymerase is a formulation of our regular BIOTAQ DNA polymerase, which contains a non-toxic and non-hazardous red dye. The red dye provides easy and quick identification of reactions to which the enzyme has been added, and facilitates the confirmation of complete mixing. When the reaction is complete, a sample of the reaction mix can be loaded directly onto the agarose gel without the need for loading buffer, since the mix is of sufficiently high density to sink to the bottom of the gel. The red dye migrates towards the positive electrode, thereby providing a means to monitor the progress of the electrophoresis. The red dye migrates like a 350bp fragment on a 2% agarose TAE gel (or 600bp on a 1% agarose).

The presence of the dye has no effect on routine enzymatic manipulations, although rare exceptions may occur. In order to produce a reaction of sufficient density to allow for the direct loading of a sample onto a gel, we recommend using a minimum of 1.5 Units per 50μl reaction.

The specificity and performance of BIOTAQ Red can be further improved with the use of 2x PolyMate Additive which is designed for GC or AT-rich DNA, "dirty" templates or sequences with a high level of secondary structure.


Notes

BIOTAQ is a trademark of Bioline.


Components

Reagent

100 Units

500 Units

2500 Units

BIOTAQ Red DNA Polymerase

1 x 20µl

1 x 100µl

5 x 100µl

10x NH4 Reaction Buffer

1 x 1.2ml

2 x 1.2ml

10 x 1.2ml

50mM MgCl2 Solution

1 x 1.2ml

1 x 1.2ml

5 x 1.2ml


Concentration

1u/µl


Storage & Stability

All kit 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 kit is retained until the expiry date indicated on the outer box label.


Shipping conditions

On Dry Ice or Blue Ice.


Certificates of Analysis (COAs)

Certificates of Analysis for BIOTAQ™ Red DNA Polymerase - Discontinued are grouped by catalogue number and then listed by batch number. Larger pack sizes are listed first. To locate the COA for your product, first find the catalogue number and then the required batch number.

View all 3 COAs for BIOTAQ™ Red DNA Polymerase - Discontinued →


Frequently asked questions

    1. Which polymerase do I need for my specific application?
    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 selection guide.

    2. What are the storage conditions and stabilities of Bioline polymerases?
    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.

    3. I am having problems optimizing my PCR, what would you recommend?
    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  For difficult templates (AT and GC rich).
     Enzyme concentration too low – increase the amount enzyme in 0.5U increments.
     Magnesium concentration too low – increase concentration in 0.25mM increments with a starting  concentration of 1.75mM.
     Primer concentration not optimised. Titrate primer concentration (0.3-1µM); 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 Hi-Spec Additive or 2x PolyMate (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 0.5U increments.
     Extension time too long. Reduce extension time in 0.5-1 minute increments.


    As an alternative for quick and simple PCR optimization, we recommend using SureBand PCR Optimization kit.

    4. What do the terms yield, fidelity, processivity and specificity actually mean?
    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.

    5. What is a proofreading polymerase?
    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.

    6. What are the error rates of Bioline’s DNA Polymerases?
    Please see the following table for the appropriate error rates:

    BIOTAQ, BIOTAQ Red, IMMOLASE,    Mango Taq
     1.1 x 10-4 base substitutions/bp (Tindall and Kunkel, 1988)
     2.4 x 10-5 frameshift mutations/bp (Tindall and Kunkel, 1988)
     2.1 x 10-4 errors/bp (Keohavang and Thilly, 1989)
     7.2 x 10-5 errors/bp (Ling et al., 1991)
     8.9 x 10-5 errors/bp (Cariello et al., 1991)
     2.0 x 10-5 errors/bp (Lundberg et al., 1991)
     1.1 x 10-4 errors/bp (Barnes, 1992)
    ACCUZYME, ACCUZYME Red, AccuSure  1.6 x 10-6 errors/base (Lundberg et al., 1991).

    7. How is one unit of activity of a polymerase defined?
    One unit is defined as the amount of enzyme that incorporates 10nmoles of dNTPs into acid- insoluble form in 30 minutes at 72°C.

    8. What is the extension rate of BIOTAQ/Red DNA Polymerase?
    BIOTAQ DNA Polymerase is a fast enzyme, which will extend fragments at 15-30s/Kb, dependant on template amplified.

    9. Is the product obtained using BIOTAQ/Red DNA Polymerase suitable for use in TA cloning?
    Yes, BIOTAQ DNA Polymerase does not possess 3’ to 5’ exonuclease activity, and therefore leaves an A-overhang at the 3’ end, thus making the PCR product suitable for integration into TA cloning vectors.

    10. What is the maximum length BIOTAQ/Red DNA Polymerase will amplify?
    BIOTAQ will comfortably amplify fragments of up to 5Kb on Genomic templates.

    11. What is the ideal extension temperature of BIOTAQ/Red DNA Polymerase?
    BIOTAQ DNA Polymerase will extend between 50-80°C, however optimal extension will occur at 72°C.

    12. Will the dye in BIOTAQ Red interfere with any applications carried out post-PCR?
    No, the dye in BIOTAQ Red is completely inert, and as such will not interfere with any downstream applications. If you are concerned about these dyes interfering in your specific applications, we would recommend the cleanup of your samples using SureClean Plus.

    13. What are the concentrations of BIOTAQ and BIOTAQ Red DNA Polymerase?
    BIOTAQ DNA Polymerase is supplied at 5u/µl, whereas BIOTAQ Red DNA Polymerase is supplied at 1u/µl.



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