DyNAzyme™ EXT DNA Polymerase

Enzyme mix for standard, difficult and long range PCR

Description

DyNAzyme™ EXT DNA Polymerase is a versatile and easy-to-use enzyme especially suited for handling demanding reaction conditions without time-consuming optimization. It is an optimized mixture of DyNAzyme™ II DNA Polymerase and a proofreading enzyme. DyNAzyme EXT DNA Polymerase offers exceptional performance that Taq-based mixtures cannot match.

Applications

Difficult templates
Templates with GC-rich or long complementary areas can be difficult to amplify. They possess strong secondary structures that resist denaturation and prevent primer annealing. DyNAzyme EXT DNA Polymerase can be used with different PCR additives, such as DMSO, formamide, glycerol and betaine, which make the denaturation easier by relaxing the template DNA. In the presence of DMSO, DyNAzyme EXT DNA Polymerase has better thermostability than Taq-based mixtures. Therefore, excellent results are obtained when amplifying difficult templates.

Long PCR
DyNAzyme EXT DNA Polymerase is an optimal and easy-to-use enzyme for long PCR. Fragments up to 40 kb have been amplified with the enzyme.

Amplification of 0.5 kb- 40 kb template using DyNAzyme EXT DNA Polymerase (1 PowerPoint slide)

Cloning
The improved fidelity and robustness of DyNAzyme EXT DNA Polymerase make it a successful enzyme for PCR cloning. DyNAzyme EXT DNA Polymerase is capable of adding a non-templated adenine residue at the 3’ end of a DNA fragment. PCR products produced with DyNAzyme EXT DNA Polymerase can be used in either TA cloning or blunt cloning.

Versatility in blunt vs. TA-cloning (3 PowerPoint slides)

Applications that require detergent-free buffers
DHPLC and microarray. A detergent-free reaction buffer is available for DyNAzyme EXT DNA Polymerase. PCR products used for DHPLC analysis are recommended to be free of detergents. Detergents can cause foaming when PCR products are spotted on microarray slides.
Buffers for DNA Polymerases

Advantages

• Ideal for difficult templates, such as GC- rich or looped sequences
• Improved fidelity compared to standard PCR enzymes
• Generates fragments up to 40 kb in long PCR reactions
• High yields, robust reactions with standard templates
• More thermostable than Taq

Contents

DyNAzyme EXT DNA Polymerase is supplied with DyNAzyme™ EXT Optimized Buffer, 10x Mg²⁺-free DyNAzyme™ EXT Buffer, 50 mM MgCl₂-solution and DMSO. 1x 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. 1x 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, stabilizers, 200 µg/ml BSA and 50 % glycerol.

Storage and stability

Recommended storage temperature is -20 °C. Stable for one year from the assay date.

Ordering information

DyNAzyme™ EXT DNA Polymerase
F-505S	200 U (1 U/µl)
F-505L	1 000 U (1 U/µl)