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As shown in Figures 2 and 3 Lucigen’s EconoTaq DNA Polymerase performs as well as, or better than, more expensive Taq preparations from several other suppliers in routine PCR. EconoTaq is as effective in DNA amplification as another standard PCR enzyme, Tfl DNA polymerase (Figure 4). In this case, the background of non-specific amplification was much lower with EconoTaq (compare “+”and “–“ lanes for EconoTaq and Tfl in Figure 4). EconoTaq DNA Polymerase also offers high lot-to-lot reproducibility and reliability (Figures 2 and 4)
Figure 1. High purity of EconoTaq DNA Polymerase (SDS PAGE). Lane 1, broad range molecular weight markers; Lane 2, Lucigen EconoTaq DNA Polymerase
Figure 2. Taq DNA polymerase from Promega and New England Biolabs were compared to Lucigen’s EconoTaq DNA Polymerase (2 different lots) in amplifying the ampicillin gene (0.8 kb) in a pUC19 vector. (–), no DNA. (+), DNA added (40 ng). MW, 1 kb ladder.
Figure 3. EconoTaq vs. AmpliTaq® (Applied Biosystems) DNA polymerase in genotyping. All PCR reactions were performed in a RoboCycler 96 (Stratagene). Hip1 genotyping was performed using the following PCR conditions: 94°C for 1min, 35 cycles of 94°C for 30sec, 62°C for 60sec, 72°C for 90sec, and 72°C for 7min. Shh, Cdo and Gas1 genotyping were performed using the following PCR conditions: 94°C for 2min, 35 cycles of 94°C for 60sec, 65°C for 60sec, 72°C for 90sec, and 72°C for 7min. All PCR reactions contained final concentrations of 1 µM of each primer, 200 µM dNTPs, 1X cresol red loading dye, and 1U of the indicated Taq polymerase. Sequences for all PCR primers have been previously published (references available).
Data courtesy of Dr. Benjamin Allen, Dept. Molecular & Cellular Biology, Harvard University.
Figure 4. PCR amplification was performed under standard conditions using three different lots of EconoTaq DNA Polymerase and buffer, or duplicate reactions with Tfl DNA polymerase and buffer (Promega). Reactions contained primers specific for the 16S ribosomal RNA gene, with Bacillus genomic DNA (+) or no DNA (-) as a template (1450 bp product expected).
Some applications in which Lucigen's EconoTaq DNA Polymerase can be used may be covered by patents issued and applicable in the United States and certain other countries. Because purchase of this product does not include a license to perform any patented application, users of this product may be required to obtain a patent license depending upon the particular application in which the product is used. The PCR process is the subject of European Patent Nos. 201,184 and 200,262 owned by Hoffman-LaRoche. Those patents expired on March 28, 2006. The corresponding PCR process patents in the United States expired on March 29, 2005. It is the sole responsibility of the buyer to ensure that use of the product does not infringe the patent rights of third parties.
5 units/µl. One unit catalyzes the incorporation of 10 nmol of dNTP into acid-insoluble material in 30 minutes at 70°C in 50 mM Tris-HCl (pH 9.0), 50 mM NaCl, 5 mM MgCl2, 200 µM dGTP, dATP, dTTP dCTP (a mix of unlabeled and [33P] dCTP), 10 µg of activated calf thymus DNA, and 0.1 mg/ml BSA.
10 mM Tris-HCl (pH 7.5), 100 mM KCl, 0.1% Triton X-100, 0.1 mM EDTA, 1mM DTT, and 50% glycerol.
10X Reaction Buffer
100mM Tris-HCl (pH 9.0), 500 mM KCl, 1% Triton X-100, and with or without 15mM MgCl2.
EconoTaq DNA Polymerase is tested in DNA amplification using a variety of templates and primers.
One unit of EconoTaq DNA Polymerase catalyzes the incorporation of 10 nmoles of dNTP into acid-insoluble material in 30 minutes at 70°C in 50 mM Tris-HCl (pH 9.0), 50 mM NaCl, 5 mM MgCl2, 200 µM dGTP, dATP, dTTP, dCTP (a mix of unlabeled and [33P]dCTP), 10 µg Activated Calf Thymus DNA, and 0.1 mg/ml BSA.
Absence of Endonuclease or Nicking Activity
Incubation of 10 U of EconoTaq DNA Polymerase with 1 µg of supercoiled pBR322 DNA for 16 hours at 70°C results in no detectable conversion to relaxed or linear forms detectable by agarose gel electrophoresis.
Absence of Exonuclease Activity
Incubation of 10 U of EconoTaq DNA Polymerase with 1 µg of HindIII-cut lambda DNA for 16 hours at 70°C resulted in no smearing of bands on agarose gels.
EconoTaq DNA Polymerase is >99% pure as determined by SDS PAGE. There is no detectable DNA contamination.