BIO-X-ACT™ Short Mix is a complete, ready-to-use, 2x reaction mix that enables PCR assays to be performed on problematic templates through the simple addition of water, template and primers.
- High performance - for templates that fail with standard Taq DNA Polymerases
- For problematic templates - works with high GC content, dirty templates, or difficult melting profiles
- Ideal for short amplicons - amplifies genomic fragments up to 3 kb
- Convenient - pre-mixed, pre-optimized 2x solution
- Ready to use - reduces risk of contamination
BIO-X-ACT™ Short Mix is a complete, ready-to-use, 2x reaction mix which enables PCR assays on problematic templates, simply by adding water, template and primers.
BIO-X-ACT Short is recommended for short genomic DNA fragments up to 3 kb and Lambda DNA up to 5 kb.
To achieve optimal reaction conditions, the BIO-X-ACT Short Mix contains BIO-X-ACT™ Short DNA Polymerase, MgCl2, ultra-pure dNTPs manufactured by Bioline, as well as additives. The mix has been optimized for a wide variety of templates and an additional 50 mM MgCl2 solution is included, should fine-tuning be required.
BIO-X-ACT Short Mix reduces the time needed to set up reactions, thereby minimizing the risk of contamination. Greater reproducibility is ensured through a reduction in the number of pipetting steps that can introduce errors.
- Suitable for cloning
- Short amplicons
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Certification of Analysis (COAs)
BIO-X-ACT Short Mix
10 x 1.25 mL
50 mM MgCl2 Solution
- BIO-25026: 500 x 50 µL Reactions: 10 x 1.25 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.
Bioline's mixes contain magnesium chloride at the following concentrations (please see table below). An additional tube of 50 mM MgCl2 solution is supplied with each mix to facilitate further optimization if required.
|Bioline Mix||Final Magnesium Concentration|
|ACCUZYME Mix||2.0 mM.|
|BioMix / BioMix Red||2.5 mM.|
|ImmoMix / ImmoMix Red||3.0 mM.|
|BIO-X-ACT Short Mix||2.0 mM.|
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.|