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DF Taq DNA polymerase is a thermostable DNA polymerase manufactured with the highest standards. It is certified as DNA free (DF) as well as RNase and DNAse free, that guarantee reproducible and efficient PCR reactions.
Please contact our exclusive distributor, Lab Supplies Scientific
DF Taq DNA polymerase is a thermostable DNA polymerase manufactured with the highest standards. It is certified as DNA free (DF) as well as RNase and DNAse free, that guarantee reproducible and efficient PCR reactions.
Taq DNA Polymerase is a thermostable enzyme that catalyzes 5’→3′ synthesis of DNA. The enzyme has no detectable 3’→5′ proofreading exonuclease activity but possesses low 5’→3′ exonuclease activity. DF Taq DNA Polymerase has high processivity up to 5 kb, while it can reliably amplify a substrate as low as 6-12 copies per reaction e.g. 16 S rRNA fragment is consistently detected by PCR containing 1.25 units of DF Taq DNA polymerase in reactions supplied with 10 fg of E.coli genomic DNA.
Source:
The Enzyme is purified from an E. coli strain carrying a plasmid with Taq DNA polymerase gene from Thermus aquaticus YT-1.
Unit definition:
One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmoles of dNTPs into acid insoluble material in 30 minutes at 72oC.
Reagents supplied:
| PD010S kit | ||
| Cat No | Description | Tubes No. |
| PD011S | DF Taq DNA polymerase, 5u/μl, 1000u, 0.2 ml
(50 mM Tris-HCl (pH 7.9 @ 25oC), 50 mM KCl, 0.1 mM EDTA, 50% glycerol, 0.5% IGEPAL, 1 mM DTT, 0.5% Tween-20) |
1 |
| BR006-1.5 | 10x Taq DNA polymerase Buffer with 15mM MgCl2
(100 mM Tris-HCl (pH 8.3), 500 mM KCl, 15mM MgCl2) |
1 |
| BR007-1.5 | 10x Taq DNA polymerase Buffer w/o MgCl2
(100 mM Tris-HCl (pH 8.3), 500 mM KCl) |
1 |
| BR008-1.5 | 25mM MgCl2 | 1 |
PCR Guidelines:
PCR recommended reaction:
| Components | 25μl assay | Final Concentration/
Quantity (25μl assay) |
50μl assay | Final Concentration/
Quantity (50μl assay) |
| 10x Taq pol. Buffer with 15mM MgCl2 | 2.5μl | 1x | 5μl | 1x |
| dNTP mix 10mM each | 0.5μl | 0.2 mM each | 1μl | 0.2 mM each |
| 10μΜ forward primer | 0.5μl | 0.2-0.5 μM | 2.5μl | 0.2-0.5 μM |
| 10μΜ reverse primer | 0.5μl | 0.2-0.5 μM | 2.5μl | 0.2-0.5 μM |
| Template DNA | Variable | 10 fg-500 ng* | Variable | 100 fg-500 ng* |
| DF Taq DNA pol. (5u/μl) | 0.25μl | 1.25 u/25 μl reaction
(0.05 u/μl) |
0.25-0.5μl | 1.25-2.5 u/50 μl reaction
(0.025-0.05 u/μl) |
| Sterile ultrapure water | Up to 25 μl | Up to 50 μl |
* Substrate quantity is mostly dependent on the complexity and the purity of substrate and should be defined after testing for each primer set/substrate combination. For highly pure DNA substrates (OD260/OD280 ~ 2 and OD260/OD230~ 2,2) dissolved in water for molecular biology, 5-10 copies of specific target substrate are enough for end product gel visualization.
PCR recommended conditions:
| Step | Temperature | Time |
| Initial denaturation | 95oC | 5 min |
| 25-35 cycles | ||
| Denaturation | 95oC | 30sec |
| Annealing | 45-68oC* | 20sec |
| Extension | 72oC | 1min/kb |
| Final extension | 72oC | 5 min |
| Hold | 4oC | Indefinitely |
*Annealing temperature depends on primers’ Tm
Functional Quality Control:
DF Taq DNA Polymerase is tested for performance in the polymerase chain reaction (PCR) using 1.25 units of enzyme to amplify:
Other Quality Controls:
Certified as DNase-RNase free using fluorescently labeled RNase and DNase probes. DNA free contamination is tested extensively through PCR for the detection of E.coli 16S ribosomal DNA. More details concerning quality controls could be found in respective Certificate of analysis.
Shipping
Shipped on blue ice or dry ice
Storage conditions
Store at -20οC ± 5οC
Shelf life:
24 months post production
Summary of the Best-Performing Characteristics of Our Taq DNA polymerase
Sensitivity and DNA-Free assay. Standard 25 μl PCR reactions were set either with EQ DF Taq or two other standard PCR polymerases from two different competitors. Reactions were supplied with either no template or 10, 100 and 1000 fg of E. coli DNA. 0.5 u of each Taq DNA pol has been added per reaction.
High processivity
Processivity assay.50 μl PCR reactions were set with either DF Taq from EQ or a competitor’s standard Taq DNA pol, to amplify a 5 kbp amplicon from λ DNA. One third of each reaction was analyzed in a standard 0.8X agarose gel prepared with our in-house Agarose (cat number: AG001-500). Marker: λ/Sty I
DF Taq DNA Polymerase is a thermostable DNA polymerase for PCR applications. "DF" stands for DNA Free, meaning the enzyme is certified to be free of DNA contamination, as well as RNase and DNase free, ensuring reproducible and efficient PCR reactions.
DF Taq DNA Polymerase has high processivity up to 5 kb. The functional quality control tests include amplification of a 5000-bp region of Lambda DNA, demonstrating its capability for longer amplicons.
The enzyme is highly sensitive and can reliably amplify substrates as low as 6-12 copies per reaction. For example, 16S rRNA fragments are consistently detected in PCR reactions containing just 10 fg of E.coli genomic DNA with 1.25 units of enzyme.
The kit includes two buffers: (1) 10x Taq Buffer with 15mM MgCl2 for standard reactions, and (2) 10x Taq Buffer without MgCl2 for optimization experiments. A separate 25mM MgCl2 solution is also provided for custom MgCl2 optimization.
