HotBegan™ Hot Start Green-Taq MasterMix
€0.00
In stock
SKU
P0720
Catalog Number: P0720 (5 x 1.25 mL (250 rxn)
For a Specific, Highly Efficient & Reliable Amplifications of DNA up 160 fg, with visual Tracking of DNA Migration
Information
Datasheet
For a Specific, Highly Efficient & Reliable Amplifications of DNA up 160 fg, with visual Tracking of DNA Migration
Information
Datasheet
HotBegan™ Hot Start Green-Taq Master Mix (2x) is an optimized ready-to-use solution containing HotBegan™ Hot Start Taq DNA Polymerase, dNTPs, MgCl2 and Stabilizers. It is inactive at room temperature and only requires addition of template, primers, and water.
HotBegan™ Hot Start Taq DNA Polymerase is a Taq DNA Polymerase bound to a proprietary antibody that blocks Polymerase activity until denaturation step occurs. The heat labile antibodies are rapidly inactivated by raising the temperature (4 minutes at 95-97 °C). This prevents or minimizes primer-dimer and nonspecific products.
MasterMix also contains an Agarose Loading Buffer including a including two tracking dyes (blue and yellow dye) for visual tracking of DNA migration and a dense compound to facilitate the drop-down of the samples into the well agarose gels. The blue dye (migrates with 3 to 5 kb DNA fragments in 1% agarose gel) and the yellow dye (migrates faster than 10 bp DNA fragments in 1% agarose gel).
Like the Taq polymerase, the enzyme has 5’→3’ Polymerase activity and a weak 5’→3’ exonuclease activity but no 3’→5’ exonuclease activity (proofreading). Before enzyme activation none of enzyme activities are detectable.
Advantages & Features
- Time-Saving Protocol: save time in the PCR process and in Agarose loading samples.
- Optimized for TA Cloning and GC Cloning.
- Accurate & Fast visual Tracking of DNA Migration: thanks to its composition that includes Green Dye Buffer.
- High specificity: minimizes unspecific amplification.
- Efficient: prevents or minimizes primer-dimer and nonspecific products.
- Great sensitivity: amplifies from femptograms of DNA targets.
- Inactive: at Room Temperature.
- Powerful: amplification of targets up to 5 kb.
Specifications
Assay Conditions:
25mM Tris-HCl pH9.0 at 25 °C, 50mM KCl, 2mM MgCl2, 0.1mg/mL gelatine, 200 μM of dATP, dGTP, dTTP, 100μM[α32-P]dCTP (0.05 μCi/nmol) and 12.5 μg activated salmon sperm DNA.
Unit definition:
One unit is defined as the amount of enzyme required to catalyse the incorporation of 10 nanomoles of dNTPs into acid-insoluble material in 30 minutes at 74 °C.
Includes
Includes for 250 rxn:
- 5 x 1.25 mL HotBegan™ Hot Start Green-Taq MasterMix (2x)*
- 1.5 mL 50mM MgCl2 Solution**
* Includes HotBegan™ Hot Start Taq DNA Polymerase, Green Dye buffer (2x), 0.4 mM of each dNTP, 4 mM Mg2+ and 24% Glycerol.
**Separate tube 50 mM MgCl2 solution is provided for further optimisation. In some cases, we recommend to optimize Mg2 + concentration.
Applications
- PCR fragments amplification for TA or GC Cloning.
- Design for High Throughput Applications.
- Amplification from a limited DNA template or low copy number genes.
Shipping & Storage
Shipped in: Gel pack.
Storage: -20 °C.
HotBegan™ Hot Start Taq DNA Polymerase is a Taq DNA Polymerase bound to a proprietary antibody that blocks Polymerase activity until denaturation step occurs. The heat labile antibodies are rapidly inactivated by raising the temperature (4 minutes at 95-97 °C). This prevents or minimizes primer-dimer and nonspecific products.
MasterMix also contains an Agarose Loading Buffer including a including two tracking dyes (blue and yellow dye) for visual tracking of DNA migration and a dense compound to facilitate the drop-down of the samples into the well agarose gels. The blue dye (migrates with 3 to 5 kb DNA fragments in 1% agarose gel) and the yellow dye (migrates faster than 10 bp DNA fragments in 1% agarose gel).
Like the Taq polymerase, the enzyme has 5’→3’ Polymerase activity and a weak 5’→3’ exonuclease activity but no 3’→5’ exonuclease activity (proofreading). Before enzyme activation none of enzyme activities are detectable.
Advantages & Features
- Time-Saving Protocol: save time in the PCR process and in Agarose loading samples.
- Optimized for TA Cloning and GC Cloning.
- Accurate & Fast visual Tracking of DNA Migration: thanks to its composition that includes Green Dye Buffer.
- High specificity: minimizes unspecific amplification.
- Efficient: prevents or minimizes primer-dimer and nonspecific products.
- Great sensitivity: amplifies from femptograms of DNA targets.
- Inactive: at Room Temperature.
- Powerful: amplification of targets up to 5 kb.
Specifications
Assay Conditions:
25mM Tris-HCl pH9.0 at 25 °C, 50mM KCl, 2mM MgCl2, 0.1mg/mL gelatine, 200 μM of dATP, dGTP, dTTP, 100μM[α32-P]dCTP (0.05 μCi/nmol) and 12.5 μg activated salmon sperm DNA.
Unit definition:
One unit is defined as the amount of enzyme required to catalyse the incorporation of 10 nanomoles of dNTPs into acid-insoluble material in 30 minutes at 74 °C.
Includes
Includes for 250 rxn:
- 5 x 1.25 mL HotBegan™ Hot Start Green-Taq MasterMix (2x)*
- 1.5 mL 50mM MgCl2 Solution**
* Includes HotBegan™ Hot Start Taq DNA Polymerase, Green Dye buffer (2x), 0.4 mM of each dNTP, 4 mM Mg2+ and 24% Glycerol.
**Separate tube 50 mM MgCl2 solution is provided for further optimisation. In some cases, we recommend to optimize Mg2 + concentration.
Applications
- PCR fragments amplification for TA or GC Cloning.
- Design for High Throughput Applications.
- Amplification from a limited DNA template or low copy number genes.
Shipping & Storage
Shipped in: Gel pack.
Storage: -20 °C.
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