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

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

Description: Titan HotTaq DNA Polymerase is 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. This prevents 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. Titan HotTaq DNA Polymerase using provided buffers was compared with three other market leaders and no visually detectable differences were seen (Figure 1).

Figure 1. Titan HotTaq DNA Polymerase compared with Hot Taq DNA Polymerases from three different market leaders. Parallel PCR reactions were performed following the supplliers’ recommendations to amplify the ~500bp and ~600bp fragments of the tested gene.

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 A1* (Mg²⁺, detergent free) Tris-HCl and KCl
10x Reaction Buffer A2 (Mg²⁺ free) Tris-HCl, KCl, detergent and glycerol
10x Reaction Buffer B1 (Mg²⁺, detergent free) Tris-HCl and (NH₄)₂SO₄
10x Reaction Buffer B2* (Mg²⁺ free) Tris-HCl, (NH4)2SO4 and detergent
10x Reaction Buffer 1 (Mg²⁺ free) 800 mM Tris-HCl, 200 mM (NH₄)₂SO₄, 0.2% w/v Tween 20
10x Reaction Buffer 2 (Mg²⁺ and detergent free) 800 mM Tris-HCl, 200 mM (NH₄)₂SO₄
25 mM MgCl2
10x Enhancer 1
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 1 is NOT a reaction buffer and should be used ONLY IF non-specific amplifications occur.

(* standard buffers)

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.

To place an order, please fill in our order form, and email to info@bioatlas.com or fax +372 616 6755.