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Cat. No.: T4DP-750 (for 750U)

Cat. No.: T4DP-3k (for 3KU)

Description

T4 DNA Polymerase is a template-dependent DNA polymerase that catalyzes DNA synthesis reactions from a primer-bound single-stranded DNA template in the 5'→3' direction. T4 DNA Polymerase possesses 3'→5' exonuclease activity but lacks 5'→3' exonuclease activity.

Features

T4 DNA Polymerase can be used to fill in 5' recessed ends or trim 3' overhangs due to its combined 5'→3' DNA polymerase activity and 3'→5' exonuclease activity. The 3'→5' exonuclease activity of T4 DNA Polymerase is more efficient on single-stranded DNA than on double-stranded DNA, meaning that unpaired regions in single-stranded DNA are more susceptible to digestion by T4 DNA Polymerase. The 3'→5' exonuclease activity of T4 DNA Polymerase is approximately 200 times higher than that of Klenow Fragment.

Applications

T4 DNA Polymerase can be used for the following reactions: smoothing of 5' or 3' recessed ends of DNA; probe synthesis through displacement reactions; synthesis of the second strand during site-directed mutagenesis; cloning of PCR products that do not rely on ligation reactions.

Source

This T4 DNA Polymerase is expressed in Escherichia coli, and the gene source is T4 phage.

Definition of activity

The amount of enzyme required to catalyze the incorporation of 10 nmol of deoxyribonucleotides (dNTPs) into polynucleotides in 30 minutes at 37°C is defined as one unit of activity.

Activity detection conditions

67 mM Tris-HCl (pH 8.8), 6.7 mM MgCl2, 1 mM DTT, 16.7 mM (NH4)2SO4, 0.2 mg/ml BSA, 0.033 mM of each dNTP, 0.4 MBq/ml [3H]-dTTP, 0.2 mM heat-denatured and partially digested calf thymus DNA.

Purity

Does not contain DNA endonucleases or RNase.

Enzyme storage solution

100mM potassium phosphate (pH 6.5), 1mM DTT, 50% glycerol.

10X Reaction Buffer

100mM Tris-HCl (pH 7.9 at 25°C), 500mM NaCl, 100mM MgCl2, 10mM DTT.

Inactivation or inhibition

T4 DNA polymerase can be completely inactivated by heating at 75°C for 20 minutes. Metal ion chelators can inhibit the activity of T4 DNA polymerase.

Precautions

• Due to its 3'→5' exonuclease activity, increasing the reaction temperature, excessive enzyme usage, failure to add dNTP, or prolonged reaction time can result in the removal of DNA bases at the 3' end, leading to gapped ends.
• The enzyme should be stored in an icebox or on ice during use and immediately placed at -20°C after use.
• This product is intended for scientific research by professionals only and should not be used for clinical diagnosis or treatment, food or drug purposes, or stored in regular residential areas.
• For your safety and health, please wear laboratory attire and disposable gloves when handling.

Storage

Store at -20°C.

Related: T4 DNA polymerase (exo-)

SBS Genetech is recognized as one of the global major leading industry players in DNA Polymerase by third-party market researchers. For more details, please visit DNA Polymerase Market Size To Reach USD 568.3 Million In 2030 | Rising Demand For Customized DNA Polymerase And Next-Generation DNA Sequencing Are Some Of The Key Factors Driving Market Revenue Growth, Says Reports and Data