Intact Genomics T4 DNA Polymerase has both a DNA-dependent DNA polymerase activity and a potent 3´→5´ exonuclease activity.
The purity of this enzyme is > 95% homogeneity as determined by SDS-PAGE using Coomassie® Blue staining (see figure below).
- 3´-overhang removal to form blunt ends (1, 2).
- 5´-overhang fill-in to form blunt ends (1, 2).
- Probe labeling using replacement synthesis (2).
- DNA library preparation for Next-generation sequencing.
- Ligation-independent cloning of PCR products.
- Second strand synthesis in site-directed mutagenesis (3).
- T4 DNA Polymerase
- 10x T4 DNA Polymerase Buffer
50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA,
50% Glycerol, pH 7.5 @ 25 ºC
10X T4 DNA Polymerase Reaction Buffer
500 mM Tris-HCl, 100 mM MgCl2, 50 mM dithiothreitol, pH 7.5 @ 25°C.
One unit of T4 DNA Polymerase converts 10 nmol of dNTPs into acid-insoluble material in 30 minutes at 37°C under standard assay conditions.
Inactivated by heating at 70°C for 15 min.
Quality Control Assays
T4 DNA Polymerase is free from detectable endonuclease and RNase activities.