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D0410 Sigma

DNA Polymerase T4 from Escherichia coli 71-18/pTL43W

buffered aqueous glycerol solution

Synonym: T4-DNA-Polymerase

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Properties

Related Categories 2.7.x.x Phosphorus containing groups, 2.x.x.x Transferases, Application Index, Biochemicals and Reagents, Enzyme Class Index,
grade   for molecular biology
form   buffered aqueous glycerol solution
concentration   5,000-20,000 units/mL
foreign activity   Endonuclease, none detected
shipped in   wet ice
storage temp.   −20°C

Description

Application

Applications include:
• Convert 3′ and 5′overhangs to blunt ends
• Site directed mutagenesis
• Label 3′-ends of double-stranded DNA
• Deletion of single strands for subcloning

Biochem/physiol Actions

T4 DNA polymerase lacks 5′ exonuclease activity but has very active 3′ exonuclease activity. This allows radiolabeling of DNA fragments by replacement synthesis. In this method, double-stranded DNA is first incubated briefly with T4 DNA polymerase in the absence of deoxynucleoside triphosphates. During this incubation, the exonuclease activity acts at both 3′-ends of the DNA fragment. The result is double-stranded DNA that has long 5′-single-strand extensions. DNA complementary to these single-stranded regions is synthesized by the enzyme when deoxynucleoside triphosphates are subsequently added.

Unit Definition

One unit converts 10 nmoles of deoxyribonucleoside triphosphate into acid-insoluble material in 30 min at 37 °C.

Physical form

Solution in 200 mM potassium phosphate, pH 6.5, 2 mM DTT and 50% glycerol (v/v)

Price and Availability

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
2

Documents

Certificate of Analysis

Certificate of Origin

Protocols & Articles

Related Content

Enzymes & Proteins

Application Index | Enzyme Index | Substrate Index | Inhibitor Index | Cofactor Index | Lectin Index
Keywords: Cell signaling, Diagnostic, Drug discovery, Molecular biology

Peer-Reviewed Papers
15

References

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