BIO-X-ACT™ Short DNA Polymerase

Cat No. Size List Price* Qty  
BIO-21065 500 Units
$499.00
 
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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 3kb. When using Lambda DNA as template, the best performance is achieved within the 100bp-5kb range.

Features & Benefits

  • 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 3kb
  • Flexible format - Available as a ready-to-use 2x reaction mix (BIO-X-ACT™ Short Mix)

Applications

  • Suitable for TA cloning
  • GC-rich templates
  • Crude sample PCR
  • Forensic applications

Description

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 3kb and Lambda DNA up to 5kb.

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.


Notes

BIO-X-ACT is a trademark of Bioline.


Components

 Reagent

250 Units

500 Units

BIO-X-ACT Short DNA Polymerase

62.5µl

125µl

10x OptiBuffer

1.2ml

2 x 1.2ml

50mM MgCl2 Solution

1.2ml

1.2ml

5x Hi-Spec Additive

1.2ml

1.2ml


Concentration

4u/µl


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.


Shipping conditions

On Dry Ice or Blue Ice.


One unit will incorporate 10nmoles of dNTPs in 30min at 72°C.


Certificates of Analysis (COAs)

Certificates of Analysis for BIO-X-ACT™ Short DNA Polymerase 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.

View all 9 COAs for BIO-X-ACT™ Short DNA Polymerase →


Frequently asked questions

  • Which polymerase do I need for my specific application?

      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 enzyme selection tool.

  • What are the storage conditions and stabilities of Bioline polymerases?

      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.
  • I am having problems optimizing my PCR, what would you recommend?

      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.

      Observation Recommended Solution(s)
      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.

  • What do the terms yield, fidelity, processivity and specificity actually mean?

      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.
  • What is a proofreading polymerase?

      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.
  • What are the error rates of Bioline DNA Polymerases?

      Please see the following table for the appropriate error rates:

      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).

  • How is one unit of activity of a polymerase defined?

      One unit is defined as the amount of enzyme that incorporates 10nmoles of dNTPs into acid- insoluble form in 30 minutes at 72°C.



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