2X Hi-G9 Taq PCR Master Mix
Thermostable DNA polymerases
Thermostable DNA polymerases
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- Thermostable DNA Polymerases
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- TA-Cloning kit and Related Enzymes
- Competent Cells
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- Sequencing Grade Reagents
Find support for technicalities of applications and product use
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2X Hi-G9 Taq PCR Master Mix
Features
- Easy to use for long range PCR
- For sensitive high throughput PCR
- Tolerates a wide range of genomic DNA templates
- Generates PCR products with 3′-dA overhangs
- It can amplify up to 8 kb product
Application
- Colony PCR
- Long PCR
- TA cloning
- Primer Extension
Unit definition
One unit of Hi-G9 Taq DNA Polymerase is the amount of enzyme required to incorporate 10 nmoles of deoxyribo nucleotide into DNA in 30 min at 74°C in its bufferoptimized condition.
Storage
-20°C to -10°C in a non-frost-free freezer. Guaranteed stable for 4 months when stored properly. 2X HiG9 Taq PCR Master Mix is stable for 17-20 freeze-thaw cycles when stored at -20°C.Avoid repeated freeze/thawing of reagents
Ordering Information
| Cat. No. | Product | Pack Size | Price |
| G4804 | 2X Hi-G9 Taq PCR Master Mix | 100 rxn | POR |
| G4804A | 2X Hi-G9 Taq PCR Master Mix | 250 rxn | POR |
| G4804B | 2X Hi-G9 Taq PCR Master Mix | 1000 rxn | POR |
Product Details Specifications Protocols & Manual Product Resource
Description
Codon optimized Hi G9 Taq DNA Polymerase gene of Thermus aquaticus was cloned and purified from E. coli host. The enzyme consists of a single polypeptide with a molecular weight of approximately 94 kDa. Hi G9 Taq DNA polymerase is heat-stable and will synthesize DNA at elevated temperatures from single-stranded templates in the presence of primer. It can be used for PCR of up to 8kb product.
2x Hi G9 Taq PCR Master Mix is a robust Taq master mix supplied with a buffer which is an optimized,
ready-to-use DNA Polymerases ideally suited to PCR applications from complex genomic template bacterial colonies and cDNA products. This convenient quick-load master mix formulation contains dNTPs and different additive buffer components to enhance processivity of G9 Taq Polymerase.
ready-to-use DNA Polymerases ideally suited to PCR applications from complex genomic template bacterial colonies and cDNA products. This convenient quick-load master mix formulation contains dNTPs and different additive buffer components to enhance processivity of G9 Taq Polymerase.
Enzyme amount & Packaging
The amount of enzyme will be 1U in 50 μl reaction finally. When cloning fragments amplified with Hi G9 Taq DNA Polymerase an extra adenine overhang base is incorporated at the 3’ end of the amplified PCR product. That A overhang is required to perform the TA cloning.
The Master Mix contains 10X Hi G9 Taq DNA reaction buffer containing 5mM MgCl2 and 0.4mM of each dNTP. Standard concentration of MgCl2 in final reaction condition is 2.5 mM. The latter option may be necessary to determine optimal conditions for amplification.
The amount of enzyme will be 1U in 50 μl reaction finally. When cloning fragments amplified with Hi G9 Taq DNA Polymerase an extra adenine overhang base is incorporated at the 3’ end of the amplified PCR product. That A overhang is required to perform the TA cloning.
The Master Mix contains 10X Hi G9 Taq DNA reaction buffer containing 5mM MgCl2 and 0.4mM of each dNTP. Standard concentration of MgCl2 in final reaction condition is 2.5 mM. The latter option may be necessary to determine optimal conditions for amplification.
Unit definition
One unit incorporates 10nmol of deoxy ribonucleotide into acid-insoluble product in 30 minutes at 74°C. Unit assay conditions: 25 mM TAPS (pH 9.3), 50 mM KCl, 2 mM MgCl2, 1 mM DTT, 0.2 mM dATP, dCTP, dGTP, dTTP utilizing M13mp18DNA as template.
Template information
For low complexity DNA (e.g. plasmid, lambda or BAC DNA): 2pg− 20ng per 50 μl reaction volume needed to be used. For high complexity genomic DNA, the amount of DNA template should be 20−200ng per 50 μl reaction volume. If cDNA synthesis reaction mixture is used directly as a source for the template, the volume used should not exceed 10% of the final PCR reaction volume.
Primer requirement
The recommendation for final primer concentration is 0.5 μM. If required, the primer concentration may be optimized between 0.2−1.0 μM. The results from primer Tm calculations can vary significantly depending on the method used. Always use the Tm calculator and instructions from reputed website to determine the Tm values of primers and optimal annealing temperature. If using a two-step PCR protocol, where both primer annealing and extension occur in a single step at 72°C, the primers should be designed accordingly.
Required additives
The recommended reaction conditions for GC-rich templates include 3% DMSO as a PCR additive, which aids in the denaturing of templates with high GC contents. For further optimization the amount of DMSO should be increased in 2 % increments. In some cases DMSO may also be required for supercoiled plasmids to relax for denaturation. Other PCR additives such as formamide, glycerol, and betaine are also compatible with Hi G9 Taq DNA Polymerase. If high DMSO concentration is used, the annealing temperature must be decreased, as DMSO affects the melting point of the primers. It has been reported that 10% DMSO decreases annealing temperature by 5.5–6.0°C.
The recommended reaction conditions for GC-rich templates include 3% DMSO as a PCR additive, which aids in the denaturing of templates with high GC contents. For further optimization the amount of DMSO should be increased in 2 % increments. In some cases DMSO may also be required for supercoiled plasmids to relax for denaturation. Other PCR additives such as formamide, glycerol, and betaine are also compatible with Hi G9 Taq DNA Polymerase. If high DMSO concentration is used, the annealing temperature must be decreased, as DMSO affects the melting point of the primers. It has been reported that 10% DMSO decreases annealing temperature by 5.5–6.0°C.
| Component | G4804 | G4804A | G4804B |
| 2x Hi G9 Taq PCR Master MixControl DNA Template
Control Primer mix |
100 reaction10 µl
10 µl |
250 reaction10 µl
10 µl |
1000 reaction10 µl
10 µl |
Protocol & Manuals
Some product description and more information to download our product brochure or manual datasheet
| Catalogue No. | Product Details | Datasheet | Brochure |
| G4804 | 2X Hi-G9 Taq PCR Master Mix, 100 rxn |  |
|
| G4804A | 2X Hi-G9 Taq PCR Master Mix, 250 rxn | |
|
| G4804B | 2X Hi-G9 Taq PCR Master Mix, 1000 rxn | |
|