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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 polymerase selection guide.
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.
As an alternative for quick and simple PCR optimization, we recommend using SureBand PCR Optimization kit.
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.
Please see the following table for the appropriate error rates:
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| BIOLASE, BIOLASE Red, IMMOLASE, Mango Taq, Taq 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, ACCUZYME Red, AccuSure |
1.6 x 10-6 errors/base (Lundberg et al., 1991). |
One unit is defined as the amount of enzyme that incorporates 10nmoles of dNTPs into acid- insoluble form in 30 minutes at 72°C.
The ACCU Range is made of our highest fidelity polymerases, providing 47-fold higher fidelity than Taq:
ACCUZYME DNA Polymerase: A thermostable enzyme possessing 5’-3’ DNA polymerase and 3’-5’ proofreading exonuclease activities.
ACCUZYME Red DNA Polymerase: An enzyme with the same properties as ACCUZYME, containing an inert red dye for easy identification of reactions with polymerase added.
AccuSure DNA Polymerase: A hot-start version of ACCUZYME, with the same high-fidelity but also added specificity.
The ACCU range of polymerases has an extremely fast extension rate compared to other proofreading enzymes, which are naturally slower than non-proofreading polymerases due to their ability to go back and correct nucleotide mis-incorporations. These polymerases will extend at a rate of as little as 30s/Kb, depending on the template amplified.
Extension should be carried out at 68°C for optimal results.
All three polymerases in the ACCU range will process up to 5Kb fragments.
AccuSure is simply a hot-start version of ACCUZYME, retaining all the same properties including extremely high fidelity.
AccuSure requires a 7-10 minute heat activation step at 95°C before starting the PCR reaction. This inactivation of the enzyme is achieved chemically, thus ensuring no enzyme activity at room temperature. This provides extremely high specificity in the PCR reaction.
As with all our colored polymerases, the dye contained in ACCUZYME Red is inert and will not interfere in further applications.
If you are concerned about these dyes interfering in your specific applications, we would recommend the cleanup of your samples using SureClean.
Please Note: ACCUZYME Red is provided at the same concentration as ACCUZYME, 2.5u/µl.
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