Applications:
-Hot Start PCR.
-Primer extension.
-TA cloning.

Description: 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.

Figure 1. Amplification of ~500bp and ~600bp target DNA fragments using Titan HotTaq DNA Polymerase.
1-3 – Titan HotTaq DNA Polymerase 
M – Atlas Star 1 Kb DNA Ladder

Source: Purified from an E. coli strain carrying an overproducing plasmid containing a modified gene of Thermus aquaticus DNA Polymerase.

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.

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.

Concentration: 5 units/µl

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). This solution should be used at a defined working concentration (1x, 2x or 3x solution). Enhancer is NOT a reaction buffer and should be used ONLY IF non-specific amplifications occur.

Shipping & Storage conditions: Shipping and temporary storage for up to 1 month at room temperature has no detrimental effects on the quality of this reagent. However, routine storage at -20ºC is strongly recommended.

Comments: This product is supplied for research use only.