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Enzymes for Molecular Biology

Enzymes for Molecular Biology: Enzymes for Molecular Biology

T4 Polynucleotide Kinase, Cloned

T4 Polynucleotide Kinase (PNK) catalyses the transfer of the g-phosphate from ATP to the 5'-hydroxyl of single- or double-stranded DNA, RNA, and nucleoside 3'-monophosphates

BioSearch Tech (Lucigen/Epicentre)

Catalogue No.DescriptionPack SizePriceQty
E0055-10D3T4 Polynucleotide Kinase, 2.5 mL at 10,000 U/mL, 25,000 U2.5ml POA Quantity Add to Order
P0503KT4 Polynucleotide Kinase, Cloned10U/µl 3,000U £213.00 Quantity Add to Order
P0525KT4 Polynucleotide Kinase Kit, 25,000 U at 10,000 U/mL25,000 U at 10,000 U/mL POA Quantity Add to Order

Description

T4 Polynucleotide Kinase (PNK) catalyses the transfer of the g-phosphate from ATP to the 5'-hydroxyl of single- or double-stranded DNA, RNA, and nucleoside 3'-monophosphates. The enzyme also removes the 3'-phosphate from 3'-phosphoryl polynucleotides, deoxyribonucleoside 3'-monophosphates, and deoxyribonucleoside 3', 5'-diphosphates to form a 3'-hydroxyl group.

T4 polynucleotide kinase
 

Unit Definition

Under standard conditions, one unit converts 1nmole of 32P from [g-32P]-ATP into an acid-insoluble form in 30 minutes at 37°C in 33 mM Tris-acetate, pH 7.8, 66 mM potassium acetate, 10mM magnesium acetate, 0.5mM DTT, 1.0mM ATP, and 0.1mg/ml micrococcal nuclease-treated calf thymus DNA as substrate. 

Storage Buffer

50% glycerol containing 50mM Tris-HCl, pH 7.5, 0.1 M NaCl, 0.1mM EDTA, 0.1% Triton® X-100, and 1 mM DTT. 

T4 PNK 10X Reaction Buffer

330mM Tris-acetate, pH 7.8, 660mM potassium acetate, 100mM magnesium acetate, and 5 mM DTT. A 10mM solution of ATP for non-isotopic applications is available separately. 

Quality Control

T4 PNK is tested in 5'-phosphorylation of nucleic acids and is free of detectable nuclease activities.




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Protocols

Protocols for: T4 Polynucleotide Kinase, Cloned

T4 Polynucleotide Kinase Protocol

(catalogue number P0505H / P0501K / P0503K) 

Please note: all protocols off site are the responsibility of the products supplier

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References

References

 

  1. Sambrook, J. et al. (1989) in: Molecular Cloning: A Laboratory Manual (2nd ed.), Cold Spring Harbor Laboratory Press, New York.
  2. Chaconas, G. et al. (1980) Meth. Enzymol. 65, 75.

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Notes

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Applications & Benefits

Applications

  • Labeling of 5´ termini of DNA and RNA with 32P or 33P for DNA sequencing, blot-hybridization experiments, or transcript mapping using Mung Bean Nuclease, S1 nuclease, or other nucleases.1,2
  • Phosphorylation of oligonucleotide linkers and other DNA or RNA molecules prior to ligation, or for use in ligation amplification reactions with Ampligase® Thermostable DNA Ligase.
  • Preparation of labeled DNA or RNA molecular weight markers for gel electrophoresis and chromatography.

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Related products

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