BIOLASE™ DNA Polymerase

Cat No. Size List Price* Qty  
BIO-21042 500 Units $164.00
 
BIO-21043 2500 Units $665.00
 
BIO-21066 10 000 Units $2,217.00
 
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BIOLASE™ is a highly purified thermostable DNA polymerase that offers high yield with minimal background. BIOLASE possesses 5’-3’ exonuclease activity and leaves an ‘A’ overhang that produces PCR product suitable for effective integration into TA cloning vectors.

Features & Benefits

  • Good standard Taq polymerase - ideal for setting up new procedures
  • Easy to use - designed for easy optimization of PCR applications
  • Flexible format - supplied with 2x PolyMate Additive for difficult or "dirty" templates
  • Suitable for TA cloning - leaves 'A' overhang

Applications

  • Routine PCR applications
  • TA cloning

Description

BIOLASE™ is a highly purified thermostable DNA polymerase offering high yield over a wide range of PCR template concentrations (fig. 1). BIOLASE is a robust preparation and delivers high yields with minimal background. BIOLASE possesses 5’-3’ exonuclease activity and leaves an ‘A’ overhang such that the PCR product is suitable for effective integration into TA cloning vectors.

BIOLASE is supplied with 10x NH4-based Reaction Buffer, which provides optimal conditions for most experiments. Additional MgCl2 is provided to allow reaction conditions to be adjusted to suit the template.

The specificity and performance of BIOLASE can be further improved with the use of 2x PolyMate Additive, which is designed for GC- or AT-rich DNA, “dirty” templates or sequences with a high level of secondary structure.


Notes

BIOLASE is a trademark of Bioline.

Components

Reagent

500 Units

2500 Units

10000 Units

BIOLASE DNA Polymerase

100µl

5 x 100µl

20 x 100µl

10x NH4 Reaction Buffer

2 x 1.2ml

10 x 1.2ml

40 x 1.2ml

50mM MgCl2 Solution

1.2ml

5 x 1.2ml

20 x 1.2ml


Concentration

  • 5u/µl

Storage & Stability

All components should be stored at -20°C upon receipt for optimum stability. Multiple freeze/thaw cycles should be avoided. For long-term storage, aliquoting is recommended.

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.


Certificates of Analysis (COAs)

Certificates of Analysis for BIOLASE™ 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 2 COAs for BIOLASE™ DNA Polymerase →

Citations

  1. Ciguatera: the detection of neurotoxins in carnivorous reef fish from the coast of Cameroon, West Africa
    Bienfang P., Oben B., DeFelice S., Moeller P., Huncik K., Oben P., Toonen R., Daly-Engel T., Bowen B.
  2. X-Linked syndrome of polyendocrinopathy, immune dysfunction and diarrhea maps to Xp11 23-Xq13 3
    Bennett C.L., Yoshioka R., Kiyosawa H., Barker D.F., Fain P.R., Shigeoka A.O., Chance P.F.
  3. Upregulation of neurotoxic serine proteases, prothrombin and protease-activated receptor 1 early after spinal cord injury
    Citron B.A., Smirnova I.V., Arnold P.M., Festoff B.W.
  4. An indirect role for upstream stimulatory factor in glucose-mediated induction of pyruvate kinase and S14 gene expression
    Kaytor E.N., Qian J., Towle H.C., Olson L.K.
  5. A polymorphism in the cystatin C gene is a novel risk factor for late-onset Alzheimer's disease
    Crawford F.C., Freeman M.J., Schinka J.A., Abdullah L.I., Gold M., Hartman R., Krivian K., Morris M.D., Richards D., Duara R., Anand R., Mullan M.J.

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.
  • What is the extension rate of BIOLASE DNA Polymerase?

      BIOLASE DNA Polymerase is a fast enzyme, which will extend fragments at 15-30s/Kb, dependant on template amplified.
  • Is the product obtained using BIOLASE DNA Polymerase suitable for use in TA cloning?

      Yes, BIOLASE DNA Polymerase does not possess 3' to 5' exonuclease activity, and therefore leaves an A-overhang at the 3' end, thus making the PCR product suitable for integration into TA cloning vectors.
  • What is the maximum length BIOLASE DNA Polymerase will amplify?

      BIOLASE will comfortably amplify fragments of up to 5Kb on Genomic templates.
  • What is the ideal extension temperature of BIOLASE DNA Polymerase?

      BIOLASE DNA Polymerase will extend between 50-80°C, however optimal extension will occur at 72°C.
  • What are the concentrations of BIOLASE DNA Polymerase?

      BIOLASE DNA Polymerase is supplied at 5u/µl.




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