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.
5 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).
Purified from an E. coli strain carrying an overproducing plasmid containing a modified gene of Thermus aquaticus DNA Polymerase.
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.
Enhancer is NOT a reaction buffer and should be used ONLY IF non-specific amplifications occur.
- 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 Enhancer
Additive 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).
- Hot Start PCR
- Primer extension
- TA cloning
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