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DreamTaq™ Green DNA Polymerase
Ready-to-load PCR product

#EP0711 Supplied with: 10X DreamTaq™ Green Buffer 25 mM MgCl2	200 u (5 u/µl) (for 160 reactions of 50 µl) 1.25 ml 0.6 ml
#EP0712 Supplied with: 10X DreamTaq™  Green Buffer 25 mM MgCl2	500 u (5 u/µl) (for 400 reactions of 50 µl) 2x1.25 ml 1.25 ml
#EP0713 Supplied with: 10X DreamTaq™  Green Buffer 25 mM MgCl2	5x500 u (5 u/µl) (for 2000 reactions of 50 µl) 10x1.25 ml 5x1.25 ml
#EP0714 Supplied with: 10X DreamTaq™  Green Buffer 25 mM MgCl2	20x500 u (5 u/µl) (for 8000 reactions of 50 µl) 40x1.25 ml 20x1.25 ml
Related Documents (in pdf, ~40-60 KB): Certificate of Analysis: #EP0711, #EP0712, #EP0713, #EP0714 MSDS (English) MSDS (English-USA) MSDS (German) Flyer (in pdf, 161 KB)
Troubleshooting (in pdf, 53 KB)

 

Features

• Save time - go directly from PCR to gel electrophoresis.
• High yields of PCR products with minimal optimization.
• Higher sensitivity compared to conventional Taq DNA polymerase.
• Amplification of long targets up to 6 kb from genomic DNA and up to 20 kb from viral DNA.
• Robust amplification of difficult templates.

Description
DreamTaq™ Green DNA Polymerase is a combination of DreamTaq™ DNA Polymerase and 10X DreamTaq™ Green Buffer. DreamTaq™ DNA Polymerase is an enhanced Taq polymerase optimized for high throughput PCR applications. It ensures higher sensitivity, longer PCR products and higher yields compared to conventional Taq DNA polymerase. DreamTaq™ generates PCR products with 3'-dA overhangs and does not incorporate dUTP.
The 10X DreamTaq™ Green Buffer includes a density reagent and two tracking dyes for direct loading of PCR products on a gel. The colored buffer does not interfere with PCR performance and is compatible with downstream applications such as DNA sequencing, ligation and restriction digestion. For applications that require PCR product analysis by absorbance or fluorescence excitation, we recommend using the colorless 10X DreamTaq™ Buffer (#B65) or purifying the PCR product prior to analysis. The 10X DreamTaq™ Green Buffer is optimized for robust performance in PCR and includes MgCl₂ at a concentration of 20 mM.

Applications

• Routine PCR amplification of DNA fragments up to 6 kb.
• RT-PCR.
• Genotyping.
• Generation of PCR products for TA cloning.

Concentration
5 u/µl

Quality Control
The absence of endo-, exodeoxyribonucleases and ribonucleases confirmed by appropriate quality tests. Functionally tested in PCR.

Definition of Activity Unit
One unit of the enzyme catalyzes the incorporation of 10 nmol of deoxyribonucleotides into a polynucleotide fraction (adsorbed on DE-81) in 30 min at 70°C.
Enzyme activity is assayed in the following mixture: 67 mM Tris-HCl (pH 8.8 at 25°C), 6.7 mM MgCl₂, 1 mM 2-mercaptoethanol, 50 mM NaCl, 0.1 mg/ml BSA, 0.75 mM activated calf thymus DNA, 0.2 mM of each dNTP, 0.4 MBq/ml [³H]-dTTP.

Storage Buffer
The enzyme is supplied in: 20 mM Tris-HCl (pH 8.0), 1 mM DTT, 0.1 mM EDTA, 100 mM KCl, 0.5% (v/v) Nonidet P40, 0.5% (v/v) Tween 20 and 50% (v/v) glycerol.

10X DreamTaq™ Green Buffer
A proprietary formulation which, in addition to the PCR buffer components, includes a density reagent and two tracking dyes. The density reagent allows direct loading of PCR products on a gel. The blue dye (migrates with 3-5 kb DNA fragments in 1% agarose gel) and the yellow dye (migrates faster than 10 bp DNA fragments in 1% agarose gel) are included for monitoring electrophoresis progress. The dyes have excitation peaks at 424 nm and 615 nm.
The 10X DreamTaq™ Green Buffer contains KCl and (NH₄)₂SO₄ at a ratio optimized for robust performance in PCR and includes MgCl₂ at a concentration of 20 mM.

Inhibition and Inactivation

• Inhibitors: ionic detergents (deoxycholate, sarkosyl and SDS) at concentrations higher than 0.06, 0.02 and 0.01%, respectively (1).
• Inactivated by phenol/chloroform extraction.

Figure 1. Longer PCR fragments and higher PCR yields obtained with DreamTaq™ Green DNA Polymerase compared to competing enzymes.
Amplification of PCR fragments from complex mouse and human genomic DNA using DreamTaq™ Green DNA Polymerase and other commercial enzymes that allow direct gel loading of PCR mixtures. PCR was performed according to supplier recommendations in a 50 µl reaction volume and 10 µl of each PCR mixture was loaded on the gel.
M – GeneRuler™ Express DNA Ladder
1 – 424 bp lcs gene fragment from mouse genomic DNA
2 – 956 bp 7 chromosome STS (G31656) from human genomic DNA
3 – 2537 bp tPA gene fragment from human genomic DNA
4 – 5003 bp beta-globine gene fragment from human genomic DNA

Related Products

10X DreamTaq™ Green Buffer 10X DreamTaq™ Buffer dNTPs dNTP Mixes dNTP Set CloneJET™ PCR Cloning Kit InsTAclone™ PCR Cloning Kit GeneJET™ Plasmid Miniprep Kit First Strand cDNA Synthesis Kit RevertAid™ First Strand cDNA Synthesis Kits FastRuler™ DNA Ladders GeneRuler™ DNA Ladders Genomic DNA Purification Kit Primers Water, nuclease-free

Reference

1. Weyant, R.S., et al., Effect of ionic and nonionic detergents on the Taq polymerase, BioTechniques, 9, 308-309, 1990.

Purchase of this product includes an immunity from suit under patents specified in the product insert to use only the amount purchased for the purchaser’s own internal research. No other patent rights  (such as 5' Nuclease Process patent rights) are conveyed expressly, by implication, or by estoppel.  Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA. For complete license disclaimer, see.

Updated vasario 26, 2009 12:41