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.
- 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)
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.
- Suitable for TA cloning
- GC-rich templates
- Crude sample PCR
- Forensic applications
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Certification of Analysis (COAs)
BIO-X-ACT Short DNA Polymerase
2 x 1.2 mL
50 mM MgCl2 Solution
5x Hi-Spec Additive
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.
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.
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.
|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.|