DyNAzyme™ EXT DNA Polymerase
High Fidelity Mixture for
standard, difficult and long range PCR
DyNAzyme™ EXT DNA Polymerase is a versatile and easy-to-use enzyme,
with powerful advantages for all PCR applications. DyNAzyme™ EXT is an
optimized mixture of DyNAzyme™ II DNA Polymerase and a proofreading
enzyme. It is uniquely suited for handling demanding reaction
conditions with high fidelity and without time consuming optimization.
DyNAzyme™ EXT offers exceptional performance that Taq-based
mixtures cannot match.
The fidelity and robustness of DyNAzyme™ EXT make it an ideal
enzyme for PCR cloning. DyNAzyme™ EXT can be used in a much broader
working range (pH, Mg2+, enzyme, dNTP concentrations) than
traditional cloning enzymes (e.g. Pfu), that are very difficult to use
and only function in a very narrow range of working conditions.
TA Cloning
DyNAzyme™ EXT works well both in blunt and TA cloning. It is
capable of adding non-templated adenine residue at the 3´end of a DNA
fragment. I.e. both blunt and 'overhang' PCR products are produced
during PCR with DyNAzyme™ EXT.
Long PCR
DyNAzyme™ EXT is an optimal and easy-to-use enzyme for long PCR.
Fragments up to 40 kb have been amplified with DyNAzyme™ EXT.
Difficult Templates
Some templates are difficult to amplify by PCR because they have
GC-rich or long complementary areas that easily form looped
structures. These templates possess strong secondary structures that
resist denaturation and prevent primer annealing. DyNAzyme™ EXT can be
used with different PCR additives, such as DMSO, formamide, glycerol
and betaine, that make the denaturation easier by relaxing the
template DNA. In the presence of DMSO, DyNAzyme™ EXT has better
thermostability than Taq-based mixtures. Due to this, excellent
results are obtained when amplifying difficult templates.
The fidelity of DyNAzyme™ EXT was compared with other enzymes and
enzyme mixtures using the immobilized mismatch binding protein (IMBP)
method. The lower the 0D450-value is, the better is the fidelity of
the enzyme. Enzyme mix A is optimized for high fidelity; mix B for
long PCR and mix C for long and accurate PCR.
DyNAzyme™ EXT is supplied with DyNAzyme™ EXT Optimized buffer, 10 x
Mg2+-free DyNAzyme™ EXT buffer, 50 mM MgCl2-solution
and DMSO. 1 x Optimized DyNAzyme™ EXT buffer contains: 50 mM Tris-HCl
(pH 9.0 at 25 °C), 1.5 mM MgCl2, 15 mM (NH4)2SO4
and 0.1 % Triton® X-100. 1 x Mg2+-free DyNAzyme™ EXT buffer
contains: 50 mM Tris-HCl (pH 9.0 at 25 °C), 15 mM (NH4)2SO4
and 0.1 % Triton® X-100.
Storage buffer
20 mM Tris-HCl (pH 7.4 at 25 °C), 0.1 mM EDTA, 1 mM DTT, 100 mM
KCl, 0.5 % Tween® 20, 0.5 % Nonidet P 40, 200 µg/ml BSA and 50 %
glycerol.
Storage stability
Stable for at least 1 year. Recommended storage temperature -20
°C.
