Oligonucleotide Characterization Services

The Overview of Oligonucleotide Characterization

Oligonucleotides are polymer sequences of nucleotides (RNA, DNA and their analogs) with chain lengths ranging from 20 to 80 nucleotides, synthesized on a scale of milligrams to kilograms. Although the synthesis of oligonucleotides is very efficient, the synthesis process produces a corresponding amount of residual oligonucleotide sequences and other process-related improvements and impurities. It is critical to analyze the synthesized products and determine their purity. Control analysis of the synthesized product and analysis of the finished product are useful for quality and process control in design studies. BOC Sciences' team of analytical experts provides professional oligonucleotide characterization services and efficiently analyzes the molecular weight, structure and impurities of the oligonucleotides through a variety of characterization techniques to facilitate research in the biological and medical fields.

Fig 1. Oligonucleotide characterization using liquid chromatography-mass spectrometry. (Sutton et al., 2022)

Why Oligonucleotide Characterization?

Oligonucleotides are much larger than conventional small molecules, and their preparation involves a large number of synthetic steps. Typical solid-phase synthesis in a multi-step cycle is likely to introduce impurities/invalid sequences, which makes analysis and characterization more difficult. In addition, oligonucleotides have certain specific physicochemical properties. Such as, they usually carry a large number of negative charges, phosphorylation to produce PO impurities, hydrolysis of purine bases to form depurine fragments and hydroxyl groups on the ribose, which are very difficult to be separated from the main product and the full-length product, and so the development of oligonucleotides usually relies heavily on mass spectrometry for analysis and quantification.

BOC Sciences' Characterization Techniques for Oligonucleotides

• High performance liquid chromatography (HPLC)
• Mass spectrometry (MALDI-TOF MS or ESI-MS)
• UV absorption spectroscopy
• Thermal denaturation analysis (spectrophotometry, thermal differential scanning calorimetry and thermal differential analysis)
• Nuclear magnetic resonance (NMR)

BOC Sciences' Oligonucleotide Characterization Services

BOC Sciences' years of expertise and experience are combined with the most advanced analytical equipment on the market to develop powerful analytical solutions for your unique oligonucleotide.

• Sequence Confirmation Services

BOC Sciences offers sequence confirmation services, which can be used to determine the exact sequence of an oligonucleotide to verify that the synthesized or extracted oligonucleotide matches the designed target sequence.

• Purity Analysis Services

BOC Sciences can detect impurities and purity levels in oligonucleotide samples using HPLC and CE techniques.

• Molecular Weight Determination Services

Mass spectrometry, our detection technology, can be used to determine the molecular weight of oligonucleotides to ensure that the synthesized oligonucleotide matches the designed target length.

• Melting Temperature Analysis Services

Thermal denaturation analysis can be used to analyze the melting temperature (T_m) of oligonucleotides, which is the temperature at which double-stranded DNA or RNA is transformed into single strands.

• Structural Validation

Our researchers can verify the structure of oligonucleotides using methods such as nuclear magnetic resonance (NMR) and mass spectrometry techniques (e.g., ESI-MS) to ensure that they have been synthesized or extracted correctly.

• Nucleic Acid Concentration Determination

BOC Sciences provides determination of the concentration of oligonucleotide samples to ensure that the amount of oligonucleotide used in an experiment is accurate.

BOC Sciences provides high-quality and efficient analytical characterization services for oligonucleotides, and offers technical support for related characterization methods, providing customized solutions for customers' oligonucleotide characterization. If you are interested in our services, please feel free to contact us.

Reference

1. Sutton J M, et al. Current state of oligonucleotide characterization using liquid chromatography–mass spectrometry: insight into critical issues[J]. Journal of the American Society for Mass Spectrometry, 2020, 31(9): 1775-1782.