Description
The ExcelTaq™ Hot Start II DNA Polymerase is a mixture of an aptamer-based inhibitor and a recombinant thermo-stable Taq DNA polymerase designed for preventing or minimizing non-specific DNA amplification in PCR reaction. The inactivation of polymerase is achieved by a reversible binding of the aptamer to the polymerase at temperatures below 45°C. The aptamer inhibitor releases polymerase during normal PCR cycling. The aptamer-based inhibition omits the time-consuming initial activation step required by chemically modified or antibody-based hot start polymerases.
The high specificity and sensitivity of ExcelTaq™ Hot Start II DNA Polymerase allows sensitive detection from limited amount of DNA templates, such as 1 pg of cDNA or 1 fg of plasmid DNA. With a high DNA synthesis rate and high thermo-stability, the ExcelTaq™ Hot Start II DNA Polymerase allows reactions to be set up at room temperature and is suitable for common and specialized PCR applications
Features
Aptamer-based hot start PCR
Reversible enzyme inactivation
Omits extra enzyme activation step
Convenient for room temperature PCR set-up
High yield and specificity of target amplicons
Wide range of amplicon length (up to 10 kb)
High sensitivity (as low as 1 fg of plasmid)
Applications
High specificity PCR
High-throughput PCR
Generation of PCR products for TA cloning
Routine PCR, multiplex PCR, colony PCR, and RT-PCR
Storage
-20°C for 24 months
Hot Start
ExcelTaq™ Hot Start II shows absolute amplicon when pre-incubation at 37˚C and 42˚C.
Reactions were subjected to a pre-incubation at 37˚C or 42˚C for 10 minutes before initial denaturation step. General Taq DNA polymerase generated primer dimers and failed to amplify target due to enzyme activated at 37˚C or 42˚C.
High specificity
ExcelTaq™ Hot Start II DNA Polymerase shows high specificity on amplifying target DNA.
The optimal annealing temperature of GAPDH primer set is 58˚C. Improper annealing temperature set at 52˚C may force primer-dimer formation. ExcelTaq™ Hot Start II DNA Polymerase eliminated primer-dimer and increased amounts of desired product. Taq DNA polymerase reduced target amplification due to primer-dimer formation at improper annealing temperature of 52˚C.
High sensitivity
ExcelTaq™ Hot Start II DNA Polymerase shows high sensitivity to amplify from low amount of templates.
Each set of PCR reactions contained either 1 pg, 10 pg, or 1 ng of HeLa cell cDNA as templates. ExcelTaq™ Hot Start II DNA Polymerase successfully amplified targets from lower amount of templates, in comparison with hot-start DNA polymerases from other suppliers and general Taq DNA polymerase.
High sensitivity
ExcelTaq™ Hot Start II DNA Polymerase amplifies from plasmid template amounts as low as 1 fg.
Contents
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Storage Buffer
50 mM Tris-HCl (pH 8.0), 50 mM KCl, 0.1 mM EDTA, 1 mM DTT, stabilizer, 50% (v/v) glycerol
10X HS buffer
200 mM Tris-HCl (pH 8.8 at 25°C), 100 mM KCl, 100 mM (NH4)2SO4, 20 mM MgCl2, 1% Triton X-100
Unit Definition
One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid-insoluble material in 30 minutes at 74°C.
Storage
-20°C for 24 months
Manual
Manual_TP5000_ExcelTaq™ Hot Start II DNA Polymerase
SDS
SDS_TP5000
Flyer
ExcelTaq™ Hot Start II DNA Polymerase
Recommended PCR Condition
Template | 1– 150 ng* |
Forward primer | 0.1– 0.5 µM |
Reverse primer | 0.1– 0.5 µM |
10X HS Buffer | 5µl |
dNTPs | 0.2 mM (each) |
Hot Start II DNA Polymerase | 0.25µl (1.25U) |
H2O | to50 µl |
Total volume | 50µl |
*Optimal amount of DNA template depends on the source and quality of DNA. The amount of purified plasmid templates can be even less than 1 pg.
Recommended PCR Program
Steps | Temp. | Time | Cycles |
Templatedenature | 94°C | 2 min | 1 |
Denature | 94°C | 30 sec | 25-40 |
Annealing | 50-68°C** | 30 sec | |
Extension | 72°C | 30 sec/kb | |
Final extension | 72°C | 1 min | 1 |
**Optimal PCR condition variesaccording to primers’ thermodynamic properties.
[RP1000] ExcelRT™ Reverse Transcriptase
High yield
Thermostable, up to 50°C, during first strand synthesis
High processivity, generating cDNA up to 8 kb
Reduced RNase H ribonuclease activity
[TP1000] ExcelTaq™ Taq DNA Polymerase
5"→3" DNA polymerase activity
5"→3" exonuclease activity
None detectable 3"→5" exonuclease (proofreading) activity
Generates PCR products with 3’-dA overhangs
Thermo-stable-half life is more than 40 min at 95°C
[TQ1200] ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, no ROX)
High Stability
Fast Hot Start
High Sensitivity
Low Background / High Specificity
Suitable for Fast Program
Smart Blue Contrast Dye
[TP1200] ExcelTaq™ 5X PCR Master Dye Mix
5’→3’ DNA polymerase activity
No detectable 3"→5" exonuclease (proofreading) activity
Generates PCR products with 3"-dA overhangs
High yield PCR
High reproducibility
Reduced pipetting errors
Includes tracking dye for direct loading after PCR
