DNA polymerase reagent Titan HotTaq

for researchfor PCR

Titan HotTaq DNA Polymerase (Fig. 1) is a modified Titan Taq DNA Polymerase. At ambient temperatures it is inactive, having no polymerase activity. Titan HotTaq DNA Polymerase is activated by a 15 minute incubation at 95-97°C, preventing the extension of non-specifically annealed primers and primer-dimers formed at low temperatures during PCR setup. The enzyme has 5’ to 3’ polymerisation-dependent exonuclease replacement activity but lacks 3’ to 5’ exonuclease activity. The enzyme has “extendase activity” allowing TA cloning. Concentration: 5 units/µl Unit definition: One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmol of dNTP into an acid-insoluble form in 30 min at 74°C. Source: Purified from an E. coli strain carrying an overproducing plasmid containing a modified gene of Thermus aquaticus DNA Polymerase. Quality control: The enzyme is free of nicking and priming activities, exonucleases and unspecific endonucleases. SDS/PAGE – 95 kD band, >98% pure. Activity and stability tested via thermocycling. The error rate per nucleotide per cycle is ~2.5 x 10-5; the accuracy is ~ 4 x 104. Estimated half life at 95°C is 1.5 hours. Storage & Dilution buffer: 50% glycerol (v/v), 20 mM Tris-HCl pH 8.7 at 25°C, 100 mM KCl, 0.1 mM EDTA and stabilizers. Supplied with: 10x Reaction Buffer 1 (Mg2+, detergent free) Tris-HCl and (NH4)2SO4 10x Reaction Buffer 2 (Mg2+ free) Tris-HCl, (NH4)2SO4 and detergent 25 mM MgCl2 10x Enhancer – Additive that facilitates amplification of difficult templates (e.g. GC-rich DNA templates).