HPLC Method for Analysis of Oligonucleotides 20 mer on BIST AC Column

Separation type: Bridge Ion Separation Technology, or BIST™ by SIELC Technologies

HPLC Method for Analysis of Oligonucleotides on BIST AC Column by SIELC Technologies

HPLC Method for Analysis of Oligonucleotides on BIST AC Column by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analysis of  Oligonucleotides

A 20 mer oligonucleotide is a short DNA molecule that is 20 nucleotides in length. Oligonucleotides of this length are commonly used in molecular biology research for a variety of applications, including PCR, DNA sequencing, gene expression analysis, and gene editing.

One common use of 20 mer oligonucleotides is as primers in PCR. PCR amplifies specific DNA sequences by using oligonucleotide primers that are complementary to the target sequence. The primers hybridize to the template DNA and serve as starting points for DNA polymerase to extend the DNA sequence in the direction of the primer.

Furthermore, 20 mer oligonucleotides are often used in gene editing techniques, such as CRISPR-Cas9. In this technique, a 20 mer oligonucleotide serves as a guide RNA (gRNA) to direct the Cas9 nuclease to the target DNA sequence, where it can make a precise cut and initiate DNA repair processes.

Overall, 20 mer oligonucleotides are versatile tools in molecular biology research that allow scientists to target specific DNA sequences for amplification, detection, and modification.

Using SIELC’s newly introduced BIST™ method, this oligonucleotide can be retained on a negatively-charged, cation-exchange BIST™ A column. There are two keys to this retention method: 1) a multi-charged, positive buffer, such as TMEDA formate, which acts as a bridge, linking the negatively charged dye to the negatively-charged column surface and 2) a mobile phase consisting mostly of organic solvent (such as MeCN) to minimize the formation of a solvation layer around the charged analytes. Using this new and unique analysis method, oligonucleotide can be separated, retained, and detected at 260 nm.

Please read more on oligonucleotides analysis by HPLC in our April’s 2023 newsletter.


ColumnBIST AC, 4.6 x 100 mm, 5 µm, 100 A
Mobile PhaseGradient MeCN – 40-5%, 10 min
BufferTMEDA formate pH 4.0 – 20 mM
Flow Rate1.0 ml/min
DetectionUV 260 nm


Class of CompoundsOligonucleotides
Analyzing CompoundsOligonucleotides

Application Column


Column Diameter: 4.6 mm
Column Length: 100 mm
Particle Size: 5 µm
Pore Size: 100 A

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Application Analytes:
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.