CYTIVA E79000Z

Thermo Sequenase DNA polymerase is a novel thermostable DNA polymerase that uses ddNTPs as readily as dNTP substrates at temperatures similar to those used in standard PCR. The result is:

ddNTPs are chain-elongating inhibitors of DNA polymerase, which are used in the Sanger method for DNA sequencing. Each nucleotide base of that particular type has a probability of being bonded not to a deoxynucleotide but rather a dideoxynucleotide, which ends chain elongation. This property, in addition to thermostability, makes Thermo Sequenase DNA polymerase an excellent choice for cycle sequencing where template quantity limitations preclude the use of T7 Sequenase DNA polymerase.

The properties of Thermo Sequenase also enable its use in many SNP genotyping, genotyping by sequencing, or genome sequencing applications where primer extension is employed.

DNA polymerases such as Thermo Sequenase can be incorporated into custom-made cycle sequencing kits, using our custom manufacturing facilities.

Assay Conditions: 25mM TAPS buffer, pH 9.3 (at 25°C), 50mM KCl, 2 mM MgCl2, 1mM 2-mercaptoethanol, 200µm dATP, dGTP, dTTP, 100µM [a-32P]-dCTP (0.05-0.1 Ci/mmol), and 400µg/ml activated DNA. After incubation at 74°C for 10 min 

Storage conditions: -20°C, Solution in 20 mM Tris-HCl, pH 8.5, 0.1 mM EDTA, 0.5% Tween 20 (v/v), 0.5% Nonidet P-40 (v/v), 1 mM DTT, 100 mM KCl, 50% glycerol 

Activity: 32 units/µl 

Source: E. coli clones that overproduce the protein 

Unit definition: One unit of enzyme incorporates 10 nmol of dNTPs into acid-insoluble material in 30 min at 74°C