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Tth DNA Polymerase Online Order

Tth DNA Polymerase

Licensed for PCR*
GENAXIS™ Biotechnology 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’ ½ 3’ DNA Polymerase lacking 3’ ½ 5’ exonuclease activity. Tth DNA Polymerase catalyzes the polymerization of nucleotides into duplex DNA in the 5’ ½ 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. GENAXIS™ Biotechnology Tth DNA Polymerase is delivered with 10X reaction buffer, 5X chelate buffer, MnCl2 and MgCl2 solutions.

APPLICATIONS: 

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

    5units/µl

REACTION BUFFER 10X:

    500mM Tris-Hcl pH8.8, 160mM (NH4)2SO4, 0.1% Tween 20. A tube of 50 mM MgCl2 and a tube of 10mM Mncl2 are provided separately.      

CHELATE BUFFER 5X:

    250mM Tris-HCl pH 8.8, 80 mM (NH4)2SO4, 3.75mM EGTA and 25% glycerol.
UNIT DEFINITION:

    one unit is defined as the amount of enzyme required to incorporate 10nmoles of dNTPs into acid precipitable form after 30 minutes at 70°C.

 COMMENTS:

    purified free of endonucleases ; good performance was confirmed using Lambda DNA as template (amplified fragment 12kb). For optimal experimental results, the Mn2+ and Mg2+ concentration and the enzyme concentration are key factors. Traditionally, 5 units enzyme and 1mM Mncl2 are used for RT activity in 50µl volume assay. For amplification, 2.5 units enzyme and 1.5mM Mgcl2 in a 50µl final volume are used.

 STORAGE AND DILUTION BUFFER:

    20mM Tris-HCl pH8.0, 100mM KCl, 0.2mM EDTA, 1mM DTT, 55% (v/v) glycerol and stabilizers. If necessary the enzyme can be diluted before use. Diluted enzyme can be stored at-20°C for up to six months.
STORAGE AND SHIPPING:

Store at -20°C. Shipped at room temperature.

* PCR is a technology covered by patents owned by Hoffmann-La Roche Inc. and F. Hoffmann-La Roche Ltd.

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