M.B. Enzymes GmbH

Tth DNA polymerase -Thermostable DNA polymerase with reverse polymerase activity.

Description

Tth DNA polymerase (EC 2.7.7.7) is a thermostable DNA-dependent DNA polymerase derived from Thermus thermophilus. The enzyme is also capable of catalyzing the polymerization of DNA in the range of 1-2 kb using an RNA template in the presence of MnCl₂. The ability of Tth DNA polymerase to reverse transcribe at elevated temperatures minimizes the problems encountered with strong secondary structures in RNA since they are unstable at higher reaction temperatures. Higher temperatures also result in increased specificity of primer hybridization and extension. Optimal temperature is 75°C, half-life at 95°C is 60 min. The enzyme will incorporate modified nucleotides (e.g., digoxigenin-11-dUTP and fluorescein-12-dUTP but not biotin-16-dUTP) at high rates. The enzyme has 3´ terminal deoxynucleotidyl transferase actifity which usualy results in addition of single dATP to the duplex DNA.

Unit definition

One unit of the enzyme will incorporate 10 nmol of dNTPs into an acid-insoluble form in 30 min at 72°C under standard assay conditions using a DNA template.

Reaction buffer

Reverse transcription buffer B6: (x10) : 670 mM Tris-HCl, pH 8.8, 166 mM (NH₄)_₂SO₄, 0.1% Tween 20. Supplied with 10 mM MnCl₂. Optimal concentration of MnCl₂ is 1-1,5 mM. DMSO should be added at a final conc. of 5%.

Amplification buffer B9: (x5): 125 mM Tris-HCl pH 8.9, 500 mM KCl, 0,25% Tween 20, 12,5 mM MgCl₂, 3,75 mM EGTA, 0,05% gelatine, 25% glycerol.

Storage

Store at -20°C.

Applications

RT-PCR.

Quality control

Activity, SDS-PAGE purity, absence of endonucleases and exonucleases, specific performance tests.

References

1. Rüttimann, C. et al. (1985) Eur. J. Biochem. 149, 41.

Catalog #

T12.3

T12.4

Pack size

500 U

1000 U

Tth DNA polymerase -Thermostable DNA polymerase with reverse polymerase activity.
Description	Tth DNA polymerase (EC 2.7.7.7) is a thermostable DNA-dependent DNA polymerase derived from Thermus thermophilus. The enzyme is also capable of catalyzing the polymerization of DNA in the range of 1-2 kb using an RNA template in the presence of MnCl₂. The ability of Tth DNA polymerase to reverse transcribe at elevated temperatures minimizes the problems encountered with strong secondary structures in RNA since they are unstable at higher reaction temperatures. Higher temperatures also result in increased specificity of primer hybridization and extension. Optimal temperature is 75°C, half-life at 95°C is 60 min. The enzyme will incorporate modified nucleotides (e.g., digoxigenin-11-dUTP and fluorescein-12-dUTP but not biotin-16-dUTP) at high rates. The enzyme has 3´ terminal deoxynucleotidyl transferase actifity which usualy results in addition of single dATP to the duplex DNA.
Unit definition	One unit of the enzyme will incorporate 10 nmol of dNTPs into an acid-insoluble form in 30 min at 72°C under standard assay conditions using a DNA template.
Reaction buffer	Reverse transcription buffer B6: (x10) : 670 mM Tris-HCl, pH 8.8, 166 mM (NH₄)_₂SO₄, 0.1% Tween 20. Supplied with 10 mM MnCl₂. Optimal concentration of MnCl₂ is 1-1,5 mM. DMSO should be added at a final conc. of 5%. Amplification buffer B9: (x5): 125 mM Tris-HCl pH 8.9, 500 mM KCl, 0,25% Tween 20, 12,5 mM MgCl₂, 3,75 mM EGTA, 0,05% gelatine, 25% glycerol.
Storage	Store at -20°C.
Applications	RT-PCR.
Quality control	Activity, SDS-PAGE purity, absence of endonucleases and exonucleases, specific performance tests.
References	1. Rüttimann, C. et al. (1985) Eur. J. Biochem. 149, 41.
Catalog #	T12.3	T12.4
Pack size	500 U	1000 U