PCR & DNA modifying enzymes

DF Taq DNA Polymerase Basic kit, 1000 U

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.

Reagents supplied: DF Taq DNA polymerase, 10x Taq DNA polymerase Buffer with 15mM MgCl2, 10x Taq DNA polymerase Buffer w/o MgCl2, 25mM MgCl2

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.

price includes packaging & shipment cost

Special offer: 5 x PD010S: 526.50 € (VAT not included)

(SKUs: PD010S)

117.00526.50

 For detailed product information please refer to the IFU

Product Details

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 where 10 fg of E.coli genomic DNA standard is added.

Source:

The Enzyme is purified from an E. coli strain carrying a plasmid with Taq DNA polymerase gene from Thermus aquaticus YT-1.

Reagents supplied:

PD010S kit
Cat NoDescription 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 MgCl21

PCR Guidelines:

PCR recommended reaction:

Components25μl assay Final Concentration/ Quantity (25μl assay)50μl assayFinal Concentration/ Quantity (50μl assay)
10x Taq pol. Buffer with 15mM MgCl22.5μl1x5μl1x
dNTP mix 10mM each0.5μl0.2 mM each1μl0.2 mM each
10μΜ forward primer0.5μl0.2-0.5 μM2.5μl0.2-0.5 μM
10μΜ reverse primer0.5μl0.2-0.5 μM2.5μl0.2-0.5 μM
Template DNAVariable10 fg-500 ng*Variable100 fg-500 ng*
DF Taq DNA pol. (5u/μl)0.25μl1.25 u/25 μl reaction (0.05 u/μl)0.25-0.5μl1.25-2.5 u/50 μl reaction (0.025-0.05 u/μl)
Sterile ultrapure waterUp 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:

StepTemperatureTime
Initial denaturation95oC5 min
25-35 cycles
Denaturation95oC30sec
Annealing45-68oC*20sec
Extension72oC1min/kb
   
Final extension72oC5 min
Hold4oCIndefinitely

*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:

  1. a 400-bp region of E.coli ribosomal DNA using 100 fg of E.coli gDNA as substrate in a 25μl reaction. The resulting PCR product is visualized as a single band on agarose gel.
  2. in a 50μl reaction. The resulting PCR product is visualized as a single band on agarose gel.

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:

12 months upon production

 

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