ACCUZYME™ Mix is a convenient, ready-to-go, 2x reaction mix engineered to provide high-fidelity and maximum experiment reproducibility.
Ultra-high fidelity PCR for subsequent 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.
ACCUZYME™ Mix, 2x
2 x 1.25 mL
10 x 1.25 mL
50 mM MgCl2 Solution
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.
Shipped on Dry Ice or Blue Ice
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.
|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.|
|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).|
|Bioline Mix||Magnesium Concentration.|
|BIO-X-ACT Short Mix||2.0mM.|