DF Taq DNA Polymerase Basic kit, 1000 U

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 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:

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 -20^οC ± 5^οC

Shelf life:

12 months upon production