Cat No. Size List Price* Qty  
BIO-25028 500 x 50µl Reactions
*For more information on pricing please contact us

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ACCUZYME™ Mix is a convenient, ready-to-go, 2x reaction mix engineered to provide high-fidelity and maximum experiment reproducibility.

Features & Benefits

  • High-fidelity - ideal for subsequent cloning
  • High processivity - amplifies fragments up to 5 kb
  • Pre-optimized mastermix - for enhance reproducibility


Ultra-high fidelity PCR for subsequent cloning

Blunt-end cloning


ACCUZYME™ Mix is a convenient, ready-to-use, 2x reaction mix possessing 5´-3´ and 3´-5´ proofreading exonuclease activities, with an error-rate of 3.0 x 10-6, even with demanding applications (fig. 1), resulting in blunt-ended amplicons of up to 5 kb in length.

ACCUZYME Mix dramatically reduces the time needed to set up reactions, thereby minimizing the risk of contamination. Greater experiment reproducibility is ensured by a reduction in the number of pipetting steps that can lead to pipetting errors.


ACCUZYME is a trademark of Bioline.



100 Reactions

500 Reactions

 ACCUZYME™ Mix, 2x

 2 x 1.25 mL

 10 x 1.25 mL

 50 mM MgCl2 Solution

 1.2 mL

 1.2 mL



Storage & Stability

All components are shipped on dry/blue ice and 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 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.

Shipping conditions

Shipped on Dry Ice or Blue Ice

Certificates of Analysis (COAs)

Certificates of Analysis for ACCUZYME™ Mix 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 10 COAs for ACCUZYME™ Mix →

Frequently asked questions

  • 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 enzyme selection tool.

  • 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.
  • 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 0.5 U increments.
       Magnesium concentration too low – increase concentration in 0.25 mM increments with a starting  concentration of 1.75mM.
       Primer concentration not optimised. 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.
      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.

  • 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.
  • 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.
  • What are the error rates of Bioline DNA Polymerases?

      Please see the following table for the appropriate error rates:

      BIOLASE, IMMOLASE, MangoTaq, MyTaq DNA Polymerase
       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  1.6 x 10-6 errors/base (Lundberg et al., 1991).

  • 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.
  • What are the advantages of using a 2x Mix rather than setting up a PCR reaction from scratch?

      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.
  • What is the concentration of the polymerase mixes?

      All Bioline PCR mixes come supplied at a 2x concentration, requiring the user simply to add template, primers and PCR grade water to a final concentration of 1x.
  • What is the recommended reaction volume of the polymerase mixes?

      We recommend a final reaction volume of 50µl, this requiring the use of 25 µL of the 2x master mix, and to make up to 50 µL using template, primers and PCR grade water.
  • What is the composition of the polymerase mixes?

      Whilst the exact composition of the mixes is proprietary information, all mixes are made up of Reaction Buffer, Magnesium, dNTPs, Polymerase and additives, designed to keep the polymerase stable in the mix, as well as provide the optimal conditions for PCR.
  • What is the final working concentration (at 1x) of MgCl2 in each mix?

      Bioline's mixes contain Magnesium Chloride at the following concentrations (please see table below). An additional tube of 50mM MgCl2 solution is supplied with each mix to facilitate further optimization if required.

       Bioline Mix  Magnesium Concentration.
      ACCUZYME Mix  2.0mM.
      BioMix  2.0mM.
      BioMix Red  2.0mM.
      BIO-X-ACT Short Mix  2.0mM.
      MangoMix  3.0mM.
      MyTaq  3.0mM.
      RANGER  1.5mM.

Other researchers use:

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