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.
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 3kb and Lambda DNA up to 5kb.
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 50mM 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.
BIO-X-ACT is a trademark of Bioline.
BIO-X-ACT Short Mix
2 x 1.25ml
10 x 1.25ml
50mM 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.
Certificates of Analysis for BIO-X-ACT™ Short 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 0.5U increments.|
|Magnesium concentration too low – increase concentration in 0.25mM increments with a starting concentration of 1.75mM.|
|Primer concentration not optimised. Titrate primer concentration (0.3-1µM); 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 Hi-Spec Additive or 2x PolyMate (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 0.5U 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 Long Mix||2.0mM.|