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T4 Polynucleotide Kinase (100u)

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M4101

Description

T4 Polynucleotide Kinase catalyzes the transfer of the γ-phosphate from ATP to the 5´-terminus of polynucleotides or to mononucleotides bearing a 5´-hydroxyl group. The enzyme, purified from recombinant E. coli, may be used to phosphorylate RNA, DNA and synthetic oligonucleotides prior to subsequent manipulations such as ligation.

Features - Benefits

  • May Be Heat-Inactivated: T4 Polynucleotide Kinase may be inactivated by heating at 68°C for 10 minutes.
  • Provided with 10X Reaction Buffer: 700mM Tris-HCl (pH 7.6 at 25°C), 100mM MgCl2, 50mM DTT.
  • Blue/White Cloning Qualified: Promega's blue/white cloning assay provides a higher level of quality control for enzymes used in cloning applications.
  • Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/custom/

Applications

  • 5´ end-labeling of single- or double-stranded DNA and RNA molecules for use as probes, for sequencing or for DNA-protein footprinting.
  • Phosphorylation of DNA prior to cloning.

For more information, see the Protocols & Applications Guide.

Specifications

QC Tests

Activity, DNase, RNase, endonuclease, end-labeling, blue/white cloning assay.

Source

Recombinant E. coli strain.

Storage Buffer

20mM Tris-HCl (pH 7.5), 25mM KCl, 2mM DTT, 0.1mM EDTA, 0.1μM ATP and 50% (v/v) glycerol.

Storage Conditions

Store at –20°C.

Unit Definition

One unit is defined as the amount of enzyme required to catalyze the transfer of 1nmol of phosphate to the 5´-OH end of a polynucleotide from [γ-32P]ATP in 30 minutes at 37°C. The reaction conditions are: 40mM Tris-HCl (pH 7.5 at 25°C), 10mM MgCl2, 5mM DTT, 0.1mM [γ-32P] ATP and 0.5mM 5´-OH polynucleotide end concentration.

For product intended use please see Patents & Disclaimers tab.

Resources

Use Restrictions

M4101, M4103, C1313 For Research Use Only. Not for Use in Diagnostic Procedures.

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