products | nucleic acid and protein tools | Thermostable DNA Polymerases

Nucleic Acid and Protein Tools

Thermostable DNA Polymerases:

SUPER TAQ* and SUPER TAQ-HC*

Features:

• very heat stable, extremely effective Taq requires only 0.05 units/50 µl volume reactions
  
• SUPER TAQ-HC (high concentration) available for special applications such as RAPD
  

Product Description

Taq DNA polymerase (from Thermus aquaticus expressed in E. coli is a highly processive
5'3' DNA polymerase without 3'5'exonuclease proof-reading activity. Taq polymerase can be used for amplification of DNA fragments by the polymerase chain reaction (PCR), amplification of single-copy genes from genomic DNA, for cycle-sequencing and for DNA-labeling. Our SUPER TAQ DNA polymerase is licensed for PCR and extremely effective compared to competitors'Taq polymerases. In addition,the enzyme is very heat-stable and can be used to amplify DNA fragments of up to 10 kb. However, for amplification of fragments longer than 5 kb we recommend SUPER TAQ plus, which gives better yields and consistency (see below).
SUPER TAQ is offered in two concentrations: Regular SUPER TAQ comes in 5 units/µl and is suited for general purposes. Alternatively, we offer SUPER TAQ-HC ("high concentration"), which comes in 15 units/µl. This is particularly useful for special applications such as RAPD (random amplified polymorphic DNA) for which large amounts of polymerase are required, but larger reaction volumes cannot be tolerated. SUPER TAQ is greater than 95% pure by SDS-PAGE analysis. Each batch is tested for absence of endo-and exonuclease activities and assayed for amplification of a 506 bp fragment of Thermus thermophilus RNase H gene (see below).

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SUPER TAQ plus*

Features:

• amplification of DNA fragments up to 20 kb
  
• high amplification yields with less enzyme
  
• higher sequence fidelity of amplified fragments due to proof reading activity
  

Product Description

SUPER TAQ plus* is designed for LA 1 ("long and accurate") PCR: LA-PCR is used for amplification of DNA fragments above 5 kb. To achieve higher yields and to improve polymerization accuracy, SUPER TAQ plus contains a mixture of SUPER TAQ and a second polymerase with 3'5' exonuclease proof-reading activity. The latter corrects misincorporations of nucleotides. Since higher yields of PCR products (especially of fragments above 1 kb) are produced, less enzyme per reaction is required. The same buffer as for SUPER TAQ can be used.

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MoBiTaq-K

Product Description

MoBiTaq-K works well in PCR and for primer extension up to 4 kb templates and is also suitable for cycle sequencing.The enzyme is a 5' deletion of the wild-type Thermus aquaticus DNA polymerase. The missing N-terminal portion is homologous to the 5' exonuclease region of E. coli DNA polymerase I. MoBiTaq-K is therefore similar to, yet distinct from, the Stoffel fragment and has a highly active and even more heat-stable DNA polymerase activity compared with the full-length enzyme. Repeated exposure to 99°C does not significantly diminish the enzyme activity allowing higher denaturation temperatures to destroy GC rich or secondary structures.

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Tth DNA Polymerase

Reverse transcription and PCR* amplification with one single enzyme

Product Description

MoBiTec's Tth DNA Polymerase is the recombinant form of the enzyme obtained from the thermophilic eubacterium Thermus thermophilus HB-8 expressed in E. coli. It is a highly processive 5' > 3' DNA Polymerase lacking 3' 5' exonuclease activity. Tth DNA Polymerase catalyzes the polymerization of nucleotides into duplex DNA in the 5' 3' direction in the presence of MgCl₂. Tth DNA Polymerase has a very efficient intrinsic reverse transcriptase (RT) activity in the presence of Mn²⁺ ions. This RT activity is not associated with RNase H activity. The ability of Tth DNA Polymerase to catalyze the polymerization of DNA, using a RNA template at high temperature, minimizes the problems encountered with strong secondary structures in RNA, since they are unstable at higher reaction temperatures. Tth DNA Polymerase is resistant to prolonged incubation at 95°C. High temperatures also result in increased specificity of primer hybridization and extension. MoBiTec's Tth DNA Polymerase is delivered with 10 X reaction buffer, 5 X chelate buffer, MnCl₂ and MgCl₂ solutions.

Applications

• (RT-PCR) of RNA up to 1000 bp in the presence of Mn²⁺ ions
  
• amplification of single-stranded DNA in the presence of Mg2+ ions
• primer extension
  
• labeling of DNA fragments with radionucleotides, digoxigenin or biotin
  

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Tth Inorganic Pyrophosphatase

Removes inhibiting pyrophosphates in PCR reactions - thermostable!

Product Description

Native, thermostable Tth Inorganic Pyrophosphatase is a hydrolase purified from Thermus thermophilus. Tth Inorganic Pyrophosphatase catalyzes the conversion of inorganic pyrophosphate to orthophosphate in reactions where pyrophosphate is accumulated. These include DNA synthesis and amplification where Tth Inorganic Pyrophosphatase provides enhanced polymerisation, by removing inhibiting pyrophosphates in the reaction.

Applications

• enhanced DNA synthesis and amplification
  
• longer fragments to be processed
  
• DNA sequencing and in vitro mutagenesis
  

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See also chapter 6 of our online catalog.

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