OptiKinase
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Affymetrix / USB
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Product
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OptiKinase
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Company
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Affymetrix / USB
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Price
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Pricing Info
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More Information
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View Company Product Page
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Applications
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5 phosphorylation of single-stranded oligonucleotides, both DNA and RNA5 phosphorylation of double-stranded DNAEfficient labeling of DNA without a bias toward the base at the 5 end5 phosphorylation of oligonucleotides with a 3 phosphate group5 phosphorylation of mononucleotides with a 3 phosphate group that can generate a substance (pNp) that can be ligated to the 3 end of DNA or RNA by T4 RNA Ligase
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Original Item Name
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OptiKinase
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Source/Expression System
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E. coli strain containing a modified clone of T4 Polynucleotide Kinase that has full kinase activity but lacks phosphatase activity.
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Catalog Number
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78334Y 1000 UN
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Quantity
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1000 units
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Concentration
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10 units/ul
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Description
Description:
USB OptiKinase™ is a modified version of T4 Polynucleotide Kinase (T4 PNK)(1,2,3) which provides greater 32P or 33P labeling efficiency of DNA. T4 PNK is a homotetrameric, bifunctional enzyme containing an N-terminal kinase domain which catalyzes the transfer of the γ-phosphate of ATP to the 5 hydroxyl end of polynucleotides and a C-terminal phosphatase domain which removes the 3 phosphate. In molecular biology applications, T4 PNK is traditionally used for radiolabelling the 5-ends of DNA and RNA. However, when phosphorylating DNA, T4 PNK displays a severe base bias as the labeling efficiency is variable depending on the nucleotide at the 5-end. Deoxyguanosine at the 5end is kinased with teh highest efficiency followed by dA, dT, and then dC which is kinased with the least efficiency(4). Unlike wild-type T4 PNK, OptiKinase exhibits no such base bias or discrimination (see Figs.1 and 2). Also, since OptiKinase lacks 3 phosphatase activity, it is particularly useful for labeling 3 phosphorylated mononucleotides or RNA with a 3 phosphate. Use OptiKinase for higher levels of phosphorylation and for greater consistency compared to standard T4 PNK.
Specifications/Features
Unit Definition:
One unit is the amount of enzyme required to incorporate 1 nmol of phosphate from radiolabeled ATP into DNA substrate in 30 min at 37°C.