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

HPLC Method for Analysis of DNA Oligo, 5 bases (AAAAA) on BIST A Column by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analysis of  DNA Oligo, 5 bases (AAAAA)

Synthesis of DNA Oligonucleotide (AAAAA)

1. Automated Synthesis:

• DNA oligonucleotides, including a sequence like “AAAAA”, are typically synthesized using automated synthesizers based on solid-phase synthesis.
• The process involves the sequential addition of nucleotide residues to the growing chain in a step-wise manner, starting from the 3′-end of the oligo.

1. Chemistry:

• The synthesis uses phosphoramidite chemistry, where each nucleotide to be added is in its protected phosphoramidite form.
• The process includes coupling, capping, oxidation, and deprotection steps.

1. Purification:

• After synthesis, the oligonucleotide is usually cleaved from the solid support and deprotected.
• It is then purified, commonly by methods like HPLC or PAGE (polyacrylamide gel electrophoresis), depending on the required purity level.

Using DNA Oligo (AAAAA)

The application of a DNA oligo with a sequence “AAAAA” can vary based on the context:

1. Research and Studies:

• Molecular Biology: In molecular biology, such oligos can be used as primers for PCR, sequencing, or as probes in hybridization experiments.
• Binding Studies: The poly(A) sequence may be used to study binding interactions with proteins, such as DNA-binding proteins or enzymes.

1. Medical and Diagnostic Applications:

• As part of diagnostic kits, especially in assays that require hybridization to a complementary sequence.

1. Educational Purposes:

• Demonstrating basic principles of nucleic acid chemistry and genetics.

1. Nanotechnology:

• In DNA nanotechnology, specific sequences of DNA are used to form structures and shapes at the nanoscale. A poly(A) sequence might be part of a larger structure.

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.

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