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JoVE Journal

Biochemistry

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Nonradioactive Assay to Measure Polynucleotide Phosphorylation of Small Nucleotide Substrates

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06:49 min

May 8th, 2020

May 8th, 2020

4,536 Views

0:05

Introduction

0:29

In Vitro RNA Kinase Reaction

1:27

Gel Electrophoresis

4:53

Results: Quantification of RNA Phosphorylation

6:06

Conclusion

Transcript

This method can answer key questions about the phosphorylation of the five-prime end of DNA or RNA molecules by a special class of enzymes known as polynucleotide kinase. The main advantage of this technique is it has the resolution to detect very

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This protocol describes a nonradioactive assay to measure kinase activity of polynucleotide kinases (PNKs) on small DNA and RNA substrates.

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