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Hot SequePol (5 U/µl)
Technical characteristics
Hot SequePol DNA polymerase I is a chemically modified SequePol DNA polymerase. At ambient temperatures it is inactive, having no polymerase activity. Hot SequePol DNA polymerase is fully activated by a 10-15 minutes incubation at 95-97°C. This prevents extension of nonspecifically annealed primers.
Source: Purified from an E. Coli strain carrying Thermus species DNA Polymerase overproducing plasmid.
Analysis condition: 25 mM Tris•HCl pH 9.0 at 25°C, 50mM KCl, 2 mM MgCl2, 0.1mg/ml gelatin, 200 µM each dATP, dGTP, dTTP, 100 µM [α32-P]dCTP (0.05µCi/nmol), 12.5 µg activated salmon sperm DNA
Unit definition: One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nmol of dNTP into acid-insoluble form in 30 minutes at 74ºC.
Activity: The enzyme has only a 5’ to 3’ polymerisation-dependent exonuclease replacement activity and enhanced efficiency for incorporating unconventional nucleotides (labeled ddNTPs & acyclo nucleotides).
IMPORTANT: To activate the polymerase, the incubation of at least 4-8 minutes at 95-97ºC MUST BE the first step of primer extension. For completeHOT TERMIPol® DNA polymerase activation 10-15! minutes at 95-97ºC is necessary.
Contents:
Hot SequePol DNA polymerase: 5 units/µl. 50% glycerol (v/v), 20 mM Tris•HCl pH 8.7 at 25ºC, 100 mM KCl, 0.1 mM EDTA and stabilizers;
10x reaction Buffer SP (500 mM Tris-HCl pH 9.5);
100 mM MgCl2.
In some applications it is recommended to use this same Buffer SP as 20x (1 x buffer final concentration is 25mM Tris-HCl) and MgCl2 final concentration is 5-10 mM. .
Routine storage of Reaction Buffer and 100 mM MgCl2 at -20ºC is recommended.
Shipping & Storage conditions: Shipping at room temperature has no detrimental effects on the quality of this enzyme and its components. However, routine storage at -20ºC is strongly recommended.
Safety warnings and precautions: This product and its components should be handled only by persons trained in laboratory techniques. It is advisable that suitable protective clothing, such as laboratory overalls, safety glasses and gloves be worn. Care should be taken to avoid contact with skin or eyes. In case of contact, wash immediately with water.
Some applications of this product may be protected and require a license which is not provided by the purchase of this product. US DNA recommends all users to obtain licenses as necessary.
Note: USDNA is currently working on cooperative efforts regarding patent cooperation to allow all users access to this higher-level product. Until finalized, no sales for commercial use without proof of license by the purchase will be allowed. Otherwise, all use of this product is limited to research use and exempt applications only. |
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