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Related Experiment Video

Updated: May 28, 2026

Chemical Triphosphorylation of Oligonucleotides

Chemical Triphosphorylation of Oligonucleotides

Published on: June 2, 2022

Highly parallel oligonucleotide purification and functionalization using reversible chemistry.

Kerri T York¹, Ryan C Smith, Rob Yang

¹Illumina, Inc., Advanced Research Group, San Diego, CA 92121, USA.

Nucleic Acids Research

|November 1, 2011

Summary

This summary is machine-generated.

Researchers created a cost-effective method for purifying and functionalizing 5'-labeled oligonucleotides. This technique enables large-scale production and parallel processing for diverse applications.

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Single-step Purification of Macromolecular Complexes Using RNA Attached to Biotin and a Photo-cleavable Linker

Single-step Purification of Macromolecular Complexes Using RNA Attached to Biotin and a Photo-cleavable Linker

Published on: January 3, 2019

Area of Science:

Biochemistry
Molecular Biology
Synthetic Chemistry

Background:

Oligonucleotide synthesis and modification are crucial for molecular biology and diagnostics.
Existing purification and functionalization methods can be costly and lack scalability.
High-throughput preparation of functionalized oligonucleotides is needed for advanced applications.

Purpose of the Study:

To develop a cost-effective and scalable method for purifying and functionalizing 5"-labeled oligonucleotides.
To enable parallel processing of numerous oligonucleotides, including complex pools.
To demonstrate the utility of the method for producing functionalized oligonucleotide pools.

Main Methods:

Utilized a 5"-hexa-His phase tag purification strategy.
Employed reversible reaction chemistry for tag exchange with desired functional groups.
Applied the method to large-scale (micromole to millimole) production.
Developed protocols for parallel processing of individual oligonucleotides and high-complexity pools.

Main Results:

Successfully purified and functionalized 5 -labeled oligonucleotides.
Demonstrated the preparation of 5 -biotin, 95-mer oligonucleotide pools with >40K complexity at micromole scale.
Achieved yields up to ~16% and purity levels of 90-99% for the oligonucleotide pools.
Showcased the method's suitability for parallel processing and large-scale synthesis.

Conclusions:

The developed method offers a cost-effective and highly parallel approach for oligonucleotide purification and functionalization.
This technique is suitable for large-scale production and can be adapted for various applications requiring modified oligonucleotides.
The method facilitates the creation of complex oligonucleotide libraries for diverse research and diagnostic purposes.