RubyTaq™ DNA Polymerase

Source: E. coli strain expressing a clone of Taq DNA Polymerase from
Thermus aquaticus ^(1-3).

Applications:

• Routine PCR amplification
• TA-vector cloning
• Amplification prior to in vitro transcription

Description:
RubyTaq™ DNA Polymerase contains high-quality USB Taq DNA Polymerase with two inert dyes that serve as tracking dyes in gel electrophoresis. This greatly simplifies analysis of PCR reactions because no loading dyes or compounds that increase sample density need to be added prior to gel electrophoresis. During gel electrophoresis, RubyTaq separates into 2 colors, magenta and yellow, for easy visualization (Fig. 1). The magenta dye runs in a range between 500 bp (2% gels) and 1500 bp (0.8% gels) and the yellow dye runs at less than 10 bp.

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Fig. 1. RubyTaq™ DNA Polymerase visualization during gel electrophoresis. RubyTaq in PCR does not require the use of additional loading buffer. Simply load onto an agarose gel directly after cycling. During electrophoresis, RubyTaq separates into 2 colors, magenta (runs between 500 bp [2% gels] and 1500 bp [0.8% gels]) and yellow (runs less than 10 bp). The indicated volumes of RubyTaq DNA Polymerase were run on a 1% TAE agarose gel.

Easy-to-see, Easy-to-Load
-Load directly to gel from thermocycler
-No additional loading buffer needed
-RubyTaq separates into 2 colors -- magenta and yellow -- during gel electrophoresis

Because the dyes are inert during PCR, RubyTaq DNA Polymerase provides the same outstanding performance as standard USB Taq DNA Polymerase (Fig. 2). In addition, the dyes do not affect the migration rates of DNA and have no effect on the following procedures:

• PCR with co-solvents (e.g., betaine and DMSO)
• Restriction digestion
• Automated or manual DNA sequencing
• Ligation-based or topoisomerase-based (TOPO^®) TA cloning
• ExoSAP-IT^® PCR Clean-Up

If the dyes must be removed prior to another application (e.g., in vitro transcription), use phenol/chloroform extraction and ethanol precipitation, or a commercially available dye removal purification product such as the PrepEase Dye Clean-Up Kit.

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Fig. 2. Performance of RubyTaq™ DNA Polymerase versus Taq DNA Polymerase. Single-copy targets were amplified from the indicated amounts of human genomic DNA. For the 455 bp numb amplicon, 50 pg of template represents about 30 total target copies in the reaction. T is Taq DNA Polymerase, R is RubyTaq DNA Polymerase, and M is the DNA marker.

Properties:
Activator: Mg²⁺

Purity:
Free from detectable non-specific nucleases.

Storage Buffer:
20mM Tris-HCl, pH 8.5, 1mM DTT, 0.1mM EDTA, 50mM KCI, 50% glycerol, inert dyes, and stabilizers.

Unit Definition:
One unit incorporates 10 nmol of total nucleotides into acid-insoluble material in 30 min at 74°C in a total volume of 50 µl.

Concentration:
1 unit/µl

Assay Conditions:
The reaction mixture (50 µl) contains 25mM TAPS, pH 9.3 (at 25°C), 50mM KCl, 2mM MgCl₂, 1mM 2-mercaptoethanol, 200µM dNTPs, 250 µg/ml activated salmon sperm DNA and cloned RubyTaq DNA Polymerase. After incubation at 74°C for 10 min, acid-insoluble material is determined.

Tested User Friendly™ Functional Test:
Functionally tested for PCR.

Functionally Tested 10X PCR Reaction Buffer without MgCl₂ (included, PN 71166):
100mM Tris-HCl (pH 8.6) and 500mM KCl

Functionally Tested MgCl₂ (included, PN 71167):
25mM solution

1. INNIS, M. A., MYAMBO, K. B., GELFAND, D. H. AND BROW, M. A. (1988) Proc. Natl. Acad. Sci. 85, 9436-9440.
2. LAWYER, F. C., STOFFEL, S., SAIKI, R. K., MYAMBO, K., DRUMMOND, R. AND GELFAND, D. H. (1989) J. Biol. Chem. 264, 6427-6437.
3. INNIS, M. A. AND GEFLAND, D. H. (1990) PCR Protocols: A Guide to Methods and Applications, Academic Press.

Storage:
Shipped on dry ice. Store at -20°C.