T4 DNA Polymerase

Description
T4 DNA Polymerase is a template dependent DNA polymerase that can catalyze DNA synthesis in the 5 '→ 3' direction in the presence of a single strand DNA template and its paired primers. T4 DNA Polymerase has 3 '→ 5' exonuclease activity, but does not have 5 '→ 3' exonuclease activity.
T4 DNA Polymerase has a stronger 3 '→ 5' exonuclease activity than DNA polymerase I, so it is a high fidelity DNA Polymerase; Unlike Escherichia coli DNA polymerase I, T4 DNA polymerase does not have 5 '→ 3' exonuclease activity.
T4 DNA Polymerase has 5 '→ 3' DNA polymerase activity and 3 '→ 5' DNA exonuclease activity at the same time, which can be used to fill the protruding end of 5 'end or flatten the protruding end of 3' end. The 3 '→ 5' DNA exonuclease activity of T4 DNA Polymerase is higher than that of double stranded DNA for single stranded DNA, that is, single stranded DNA is more easily digested by T4 DNA Polymerase than the unpaired part of double stranded DNA. The 3 '→ 5' exonuclease activity of T4 DNA Polymerase is about 100 times higher than that of Klenow Fragment.

Application: smoothing and cloning of 5 'or 3' protruding ends of DNA; The labeled DNA probe was synthesized by displacement reaction; Synthesis of the second chain during site directed mutagenesis; Cloning of PCR products independent of linkage reaction; The starting point of mRNA transcription was analyzed by primer extension.

Source: T4 DNA polymerase was expressed and purified from Escherichia coli, and the expression gene was T4 phage gene 43.

Unit definition: One unit is defined as the amount of enzyme that will incorporate 10nmol dNTP into acid insoluble material in 30 minutes at 37 °C

Purity: no DNA endonuclease, no RNase.

Storage Buffer: 100mM potassium phosphate (pH6.5), 1mM DTT, 50% glycerol.

10X Reaction Buffer: 100mM Tris-HCl (pH7.9 at 25°C), 500mM NaCl, 100mM MgCl2, 10mM DTT。

Inactivation or inhibition: heating at 75 °C for 20 min can fully inactivate T4 DNA polymerase; Metal ion chelators can inhibit the activity of T4 DNA polymerase.

Storage condition: - 20 °C.

Note:
Because the enzyme has the activity of 3 '→ 5' nucleic acid exonuclease, the increase of reaction temperature, the use of excessive enzyme, the absence of dNTP or the long reaction time may cause the DNA3 'terminal base to be cut off and form a concave end.
When using, the enzyme should be stored in an ice box or on an ice bath, and should be stored at - 20 °C immediately after use.
This product is only used for scientific research by professionals, and shall not be used for clinical diagnosis or treatment, food or medicine, or stored in ordinary houses.
For your safety and health, please wear lab clothes and disposable gloves.