Titan Hot Taq DNA Polymerase
Titan Hot Taq DNA Polymerase is useful for hot start PCR. It is similar than Titan Taq DNA Polymerase but it has been modified. Titan Hot Taq DNA Polymerase is inactive at ambient temperatures without polymerase activity, but it is activated at 95-97°C during 15 minute.The advantage is that in this way you prevent the extension of non-specifically annealed oligonucleotides and primer-dimers formed at room temperature during PCR setup. Titan Hot Taq DNA Polymerase has 5’ to 3’ polymerisation-dependent exonuclease replacement activity but lacks 3’ to 5’ exonuclease activity. Titan Taq DNA Polymerase has “extendase activity” which allows TA cloning. Vrisko Ltd provides a Titan Hot Taq DNA Polymerase under the product name Vrisko Titan HotTaq.
Vrisko Titan HotTaq
- Hot Start PCR.
- Primer extension.
- TA cloning.
Figure 1. Amplification of ~500bp and ~600bp target DNA fragments using Vrisko Titan Hot Taq DNA Polymerase.
1-3 – Vrisko Titan HotTaq DNA Polymerase
M – Vrisko Star 1 Kb DNA Ladder
Concentration: 5 units/µl
Unit definition: Amount of Vrisko Titan HotTaq 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-, exonucleases- and unspecific endonucleases activities. SDS/PAGE – 95 kD band, >98% pure. Activity and stability tested via thermocycling. The rate of error 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.
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: Vrisko Titan HotTaq is supplied for research use only.
|Product||Quantity||Cat. #||Data Sheet||MSDS|
|Vrisko Titan HotTaq||500 units||VP0019|
|Vrisko Titan HotTaq||1000 units||VP0020|