Destinado a amplificação de alvos difíceis e longos 9<40 kb)
|F-505L||1,000 units||1 units/μl|
|200 units||1 units/μl|
* Licensed for PCR*
* Exceptional Performance - masters demanding reaction conditions without time consuming optimization
* Excellent for long templates - amplifies up to 40 kb
* Ideal for amplifying difficult templates, such as GC-rich or looped sequences
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.
Figure 1: Thermal stabilities of DyNAzyme I, II and EXT were tested at 96°C. The half-life of DyNAzyme I is 3 h and that of DyNAzyme™ II 2.5 h. The half-life of DyNAzyme EXT is even better (3.5 h). In comparison with Taq polymerase, the thermal stabilities of all three DyNAzyme polymerases show clear differences starting from 120 minutes of incubation at 96°C. All DNA polymerases were diluted with the recommended assay buffers to 0.04 U/ µl. Diluted DNA polymerases were heated at 96°C . Samples were taken after 0, 15, 30, 60, 90, 120 and 240 minutes.
Figure 2: Thermal stabilities of DyNAzyme I, II and EXT were tested at 94°C with 5 % DMSO. The half-lives of DyNAzyme I and II are 40 minutes, and that of DyNAzyme EXT is 1 h. In comparison with Taq polymerase, all three DyNAzyme polymerases show better thermostability. When amplifying difficult templates containing GC-rich or long complementary areas, PCR additives such as DMSO are needed. DyNAzyme polymerases have better thermostability than Taq and Taq-based mixtures when DMSO is used. All DNA polymerases were diluted with the recommended assay buffers to 0.04 U/ µl. Buffers contained 5 % DMSO. Diluted DNA polymerases were heated at 94°C. Samples were taken after 0, 5, 15, 30, 60, 90 and 120 minutes.
DyNAzyme™ EXT DNA Polymerase is an optimized mixture of DyNAzyme II DNA Polymerase and a proofreading enzyme. DyNAzyme II DNA Polymerase is isolated and purified from an E. coli strain expressing the cloned DyNAzyme DNA Polymerase gene from Thermus brockianus.
* 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. Due to this 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 DyNAzyme™ EXT DNA Polymerase.
* 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 amplified with DyNAzyme EXT DNA Polymerase can be used in either TA cloning or blunt cloning. DyNAzyme EXT DNA Polymerase 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.
* DHPLC: PCR products used for DHPLC analysis are recommended to be free of detergents. A detergent-free reaction buffer is available for DyNAzyme EXT DNA Polymerase.
* Microarray: Detergents can cause foaming when PCR products are spotted on microarray slides. A detergent-free reaction buffer is available for DyNAzyme EXT DNA Polymerase.
- Improved fidelity compared to standard PCR enzymes
- Generates fragments up to 40 kb in long PCR reactions
- Ideal for difficult templates, such as GC- rich or looped sequences
- High yields, robust reactions with standard templates
- More thermostable than Taq
- Optimized DyNAzyme™ EXT Buffer (Mg2+-free buffer also supplied)
- DMSO (100 %)
- MgCl2 solution (50 mM)
- Enzyme Properties
- Polymerase Properties | Thermophilic Polymerase Characteristics
- 3´ to 5´ Exonuclease: Yes
- 5´ to 3´ Exonuclease: Yes
Reaction & Storage Conditions
- 1X Optimized DyNAzyme™ EXT Buffer (Mg2+-free buffer also supplied)
- 1X Optimized DyNAzyme™ EXT Buffer (Mg2+-free buffer also supplied):
- 50 mM Tris-HCl
- 15 mM (NH4)2SO4
- 1.5 mM MgCl2
- 0.1 % Triton X-100
- pH 9.0 @ 25°C
- 1 units/μl
- 20 mM Tris-HCl
- 100 mM KCl
- 1 mM DTT
- 0.1 mM EDTA
- 200 µg/ml BSA
- 50% Glycerol
- 0.5% Tween-20
- 0.5% Nonidet P40
- pH 7.4 @ 25°C
Stable for at least one year.
Protocols for DyNAzyme™ EXT DNA Polymerase
DyNAzyme™ EXT Buffer Pack
DyNAzyme™ EXT PCR Kit
Optimized Detergent-free DyNAzyme™ EXT Buffer
Optimized DyNAzyme™ EXT Buffer and 50 mM MgCl2
* PCR license notice: These products are sold under licensing arrangements of Finnzymes Oy with F. Hoffman-La Roche LTD. The purchase of these products is accompanied by a limited license to use them in the Polymerase Chain Reaction (PCR) process in conjunction with a thermal cycler whose use in the automated performance of the PCR process is covered by the up-front fee, either by payment to Applied Biosystems or as purchased, i.e. an authorized thermal cycler.