Description Methylamp Hot Taq DNA Polymerase is a chemically modified Methylamp Taq DNA Polymerase. At ambient temperatures it is inactive, having no polymerization activity. Methylamp Hot Taq DNA Polymerase is activated by a 15 min incubation step at 95°C. This prevents extentension of non-specifically annealed primers and primer-dimers formed at low temperatures during PCR setup. The enzyme has 5’→3’ polymerization dependent exonuclease replacement activity but lacks 3’→ 5’ exonuclease activity.

Concentration5 units/µl (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).

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

Storage & Dilution Buffer50% glycerol (v/v), 20 mM Tris-HCl pH 8.7 at 25°C, 100 mM KCl, 0.1 mM EDTA and stabilizers.

Composition

• Methylamp Hot Taq DNA Polymerase
• 10x Reaction Buffer B1 (Mg2+ , detergent free): Tris-HCl and (NH4)2SO4
• 10x Reaction Buffer B2 (Mg2+ free): Tris-HCl, (NH4)2SO4 and detergent
• 25 mM MgCl2
• 10x EnhancerAdditive that facilitates amplification of difficult templates (e.g. GC-rich DNA templates).Enhancer should be used at a defined working concentration (1x, 2x or 3x solution).

Applications

• Hot Start PCR
• Primer extension
• TA cloning

Storage Conditions 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.

Fig. 1.Amplification of ~500bp and ~600bp target DNA fragments using Methylamp Hot Taq DNA Polymerase.

1-3 – Methylamp Hot Taq DNA Polymerase

M – EpiQuik 1 Kb DNA Ladder