OneTaq Hot Start DNA Polymerase M0481S from New England Biolabs is a high‑fidelity enzyme supplied with standard, GC and high‑GC buffers for robust PCR.
OneTaq Hot Start DNA Polymerase is an optimized blend of Taq and Deep Vent® DNA polymerases with an aptamer‑based inhibitor that prevents activity below 45 °C. The enzyme is supplied at 200 units (5,000 U ml⁻¹) and can be stored at –20 °C. It includes two 5× reaction buffers (Standard and GC) and a High GC Enhancer to improve yields for GC‑rich templates.
The datasheet provides detailed buffer compositions, unit definition, recommended enzyme concentrations, Mg²⁺ requirements, and thermocycling guidelines. It also lists applications such as high‑sensitivity, high‑throughput, GC‑rich, AT‑rich, colony and long PCR (up to ~6 kb). Companion products and licensing information are included.
| Units per vial | 200 units (5,000 U ml⁻¹) |
|---|---|
| Storage temperature | -20 °C |
| Standard Reaction Buffer composition (1X) | 20 mM Tris‑HCl pH 8.9, 1.8 mM MgCl₂, 22 mM NH₄Cl, 22 mM KCl, 0.06 % IGEPAL CA‑630, 0.05 % Tween 20 |
| GC Reaction Buffer composition (1X) | 80 mM Tris‑SO₄ pH 9.2, 2 mM MgSO₄, 20 mM (NH₄)₂SO₄, 5 % Glycerol, 5 % DMSO, 0.06 % IGEPAL CA‑630, 0.05 % Tween 20 |
| High GC Enhancer composition (1X) | 10 mM Tris‑HCl pH 9.2, 25 % DMSO, 25 % Glycerol |
| Unit definition | Incorporates 15 nmol dNTP into acid‑insoluble material in 30 min at 75 °C |
| Recommended enzyme concentration | 1.25 units per 50 µl reaction (25 units ml⁻¹) |
| Mg²⁺ concentration (Standard Buffer) | 1.8 mM |
| Mg²⁺ concentration (GC Buffer) | 2.0 mM |
One unit is the amount of enzyme that will incorporate 15 nmol of dNTP into acid‑insoluble material in 30 minutes at 75 °C.
No separate activation step is required; the inhibitor releases during normal cycling.
The product includes two 5× buffers (Standard and GC) and a High GC Enhancer solution.
Store the enzyme at –20 °C.