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Related Concept Videos

DNA Isolation01:24

DNA Isolation

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
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Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...

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Related Experiment Video

Updated: May 11, 2026

Chemical Triphosphorylation of Oligonucleotides
13:19

Chemical Triphosphorylation of Oligonucleotides

Published on: June 2, 2022

A convenient and efficient purification method for chemically labeled oligonucleotides.

Jihee Hwang1, Junhee Kang, Seong Keun Kim

  • 1WCU Department of Biophysics and Chemical Biology, Seoul National University, Korea.

Biotechniques
|May 14, 2013
PubMed
Summary
This summary is machine-generated.

We developed a fast and cost-effective method to purify chemically-labeled oligonucleotides using a simple phase extraction technique. This new approach significantly reduces processing time compared to traditional methods.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Chemical Synthesis

Background:

  • Chemically labeling oligonucleotides is crucial for various molecular biology applications.
  • Conventional purification methods like ethanol precipitation and size-exclusion chromatography are time-consuming and can be inefficient.
  • There is a need for rapid, cost-effective, and scalable purification techniques for labeled oligonucleotides.

Purpose of the Study:

  • To develop an efficient, rapid, and cost-effective purification method for chemically-labeled oligonucleotides.
  • To offer an alternative to traditional purification techniques, suitable for high-throughput applications.

Main Methods:

  • Utilized the differential hydrophilic and hydrophobic properties of DNA and amine-reactive fluorophores.
  • Developed a phase extraction technique using n-butanol saturated with distilled water.
  • Separated labeled oligonucleotides (aqueous phase) from unreacted fluorophores (organic phase).

Main Results:

  • Achieved efficient purification of chemically-labeled oligonucleotides.
  • Demonstrated a significant reduction in purification time compared to ethanol precipitation and size-exclusion chromatography.
  • The method is simple, fast, and amenable to simultaneous processing of multiple samples.

Conclusions:

  • The developed n-butanol phase extraction method provides an efficient, rapid, and cost-effective way to purify chemically-labeled oligonucleotides.
  • This technique is suitable for high-throughput labeling strategies, overcoming limitations of conventional methods.
  • The simplicity and speed of this method make it a valuable tool for researchers working with labeled nucleic acids.