Diamond Taq DNA Polymerase

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Application
Polymerase Chain Reaction (PCR), DNA Amplification (DNA Amp)
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Purpose Diamond Taq® polymerase* is a highly thermostable enzyme produced and purified from recombinant Escherichia coli bacterium containing the Thermus aquaticus DNA Polymerase gene.
Brand Diamond Taq®
Specificity Diamond Taq® polymerase* is a highly thermostable enzyme produced and purified from recombinant Escherichia coli bacterium containing the Thermus aquaticus DNA Polymerase gene. The expressed enzyme catalyzes 5'→3' synthesis of DNA with no detectable 3'→5' proof reading exonuclease activity. This enzyme has the "extendase" activity allowing TA cloning.
Characteristics All the Diamond Taq® enzymes have been manufactured under fully compliant cGMP conditions, purifications and quality controls to offer you the maximum security in your assay.

Diamond Taq® family enzymes are highly thermostable polymerases produced and purified from recombinant Escherichia coli bacteria containing the Thermus aquaticus DNA Polymerase gene.

The expressed enzyme shows very good fidelity and catalyzes 5'>3' synthesis of DNA with no detectable 3'>5' exonuclease activity. The enzyme has the "extendase" activity allowing TA cloning.

Qualities:
  • Higher Sensitivity
  • Higher lot-to-lot consistency
    • Strickly monitored GMP and analytical processes ensuring lot-to-lot consistency
  • Ultra-low bioburden
    • Production in a GMP-Pharma facility leading to a guaranteed bioburden between 0 and 10 CFU/mL.
  • Ultra-low residual DNA content
    • QC-tested ensuring less than 1 fg (0.2 copy) of genomic E. coli DNA / Taq unit.
  • Lower risk of false positive results due to residual DNA contamination (bacterial & fungal).
Purity > 98 % by SDS-PAGE
Components Diamond Taq® is provided at a concentration of 5 U/μl with 10x reaction buffer and MgCl2 solution.
  • 10x Reaction buffer: 750 mM Tris-HCl, 200 mM (NH4)2SO4, 0.1 % (v/v) Tween 20 and stabilizer, pH 8.8 (at 19 °C).
  • MgCl2 solution: 25 mM MgCl2
  • Enzyme storage buffer: 20 mM Tris-HCl, 1 mM DTT, 0.1 mM EDTA, 0.1 M KCl, 0.5 % (v/v) Nonidet P40, 0.5 % (v/v) Tween 20, 50 % (v/v) glycerol and stabilizer pH 8.0 (19 °C)
ProductDetails: Biological Activity Comment
  • Performance test: PCR on λ DNA - 0.5 kb fragment positive down to 5 pg
  • Performance test: PCR on genomic DNA - 0.1 kb fragment positive down to 10 pg
  • Ribonucleases (up to 10 U, 1 h, 37 °C): Not detectable
  • Endonucleases (up to 10 U, 16 h, 65 °C): Not detectable
  • Exonucleases (up to 10 U, 16 h, 65 °C): Not detectable
  • Nicking activity (up to 10 U, 16 h, 65 °C): Not detectable
  • E. coli residual DNA: < 1 fg / Taq Unit
Unit Definition One unit is defined as the amount of enzyme that incorporates, 10nmoles of dNTPs into acid insoluble form in 30 minutes at 74 °C.
Molecular Weight MW approx. 95 kDa (SDS-PAGE)
Application Notes Diamond Taq® is particularly suited for PCR applications that require high sensitivity and ultra low level of bacterial & fungal DNA. The GMP manufacturing & purification processes minimize the risk of false positive results due to residual DNA contamination (bacterial or fungal). The enzyme is QC-tested to verify that < 1fg of genomic E. coli DNA (or 0.2 copy) is present in a standard aliquot containing 1 unit of Taq. Bioburden is guaranteed ≤ 10 CFU/mL, but is typically = 0 CFU/mL
Comment

Diamond Taq® polymerase is particularly recommended for PCR applications that require ultra low levels of bacterial & fungal DNA in Diagnostic and demanding Research fields. Visual confirmation of pipetting (inert red dye) can be included in the storage buffer, see Red Diamond Taq®.

Protocol For a 100 μL Reaction
  • Diamond Taq® Reaction Buffer (10x) 10 μL
  • MgCl2 solution 6 μL (1.5 mM)
  • Diamond Taq® 0.8 to 2.5 units
  • dNTP 200 μM each
  • Primers 0.1 nmol each
  • H2O as required
  • DNA template as required
Classical PCR protocol used for 500 bp lambda DNA amplification:
  • 95 °C (3 min)
  • 94 °C (30 sec)
  • Tm -2 °C (30 sec)
  • 72 °C (1 min/kb)
  • 72 °C (7 min)
  • 4 °C end temperature
at 25 cycles

Note: Condition will vary from reaction to reaction and may need optimization for maximal performances. Duration and temperature for denaturation and annealing steps depend on the type of cycler and primers design. We advise you to check primer design by using primer design Software.
Restrictions For Research Use only
Buffer Storage and dilution buffer:
  • 20 mM Tris-HCl
  • 1 mM DTT
  • 0.1 mM EDTA
  • 0.1 M KCl
  • 0.5 % (v/v) Nonidet P40
  • 0.5 % (v/v) Tween 20
  • 50 % (v/v) glycerol, pH 8.0 (19 °C) and stabilizer
Handling Advice Recommendation:
Homogenize Diamond Taq® solution by flipping the tube 4 to 5 times.

Magnesium: This DNA polymerase is a magnesium-dependent enzyme. We recommend increasing the magnesium concentration for long DNA fragments. Excess Mg2+ stabilizes the DNA double strand and consequently prevents complete denaturation of DNA, which reduces the extension yield. It may also stabilize spurious primer/ template annealing, thus decreasing specificity
Storage -20 °C
Storage Comment 24 months (at -20 °C) from date of manufacture
Expiry Date 24 months