Oligonucleotides

Oligonucleotides are short, synthetically produced DNA or RNA sequences consisting of just a few nucleotides. They play an important role in molecular biology and medicine, particularly in genetic research, diagnostics and therapy. In medicine, oligonucleotides are mainly used as therapeutic agents to influence specific genes. These include antisense oligonucleotides, which suppress gene expression, and siRNA (small interfering RNA), which specifically degrade mRNA in order to prevent the synthesis of disease-causing proteins. These technologies offer promising approaches for the treatment of genetic diseases, cancer and viral infections.

 

Analysing oligonucleotides is crucial as it ensures the quality, purity and efficacy of these molecules in biomedical applications. Precise analyses make it possible to detect sequence errors, check the functionality and stability of oligonucleotides and ensure that they have the desired biological effect. Particularly in therapeutic applications, such as antisense oligonucleotides or siRNA, precise characterisation is necessary to avoid undesirable side effects and ineffective treatment. Methods such as mass spectrometry, HPLC and sequencing play a central role here.

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Technical Data

Differences between Nucleotides, Oligonucleotides and DNA/RNA

The differences between oligonucleotide, nucleotide and DNA/RNA analysis using HPLC lie in the target molecules and the analysis methods:

 

  • Oligonucleotide analysis: short chains of nucleotides (oligonucleotides) are analysed here. Reversed-phase HPLC (RP-HPLC) is often used to separate different oligonucleotides based on their length and chemical modifications.
  • Nucleotide analysis: Individual nucleotides (ATP, GTP, CTP, UTP etc.) are analysed. Here, anion exchange HPLC is often used to separate the nucleotides based on their charge and size.
  • DNA/RNA analysis: When analysing larger DNA or RNA molecules, HPLC is used for purity testing and quantification. Specialised columns and methods such as size exclusion HPLC or affinity chromatography can be used to separate intact nucleic acids or their fragments.

Applications

Oligonucleotide separation with good peak shape with AMT HALO Oligo C18

  • Separation of oligonucleotides up to 60 bases (60 mer) with good peak shape

Peak Identities

1. 10 mer  2. 15 mer  3. 20 mer  4. 25 mer  5. 30 mer  6. 40 mer  7. 50 mer  8. 60 mer  * Tris HCl/EDTA

Column

AMT: HALO 120Å OLIGO C18, 2.7µm, 50x2.1mm

Competitor: OLIGO 130Å C18, 1.7µm, 50x2.1mm

Downloads

Manufacturer overview




Hamilton

HPLC - Reversed Phase:

  • PRP-C18, 5µm




Nouryon - Kromasil

UHPLC - Reversed Phase:

  • Kromasil 100 Phenyl, 2.5µm
  • Kromasil 100 C4, 2.5µm
  • Kromasil 100 C18, 2.5µm

Osaka Soda

UHPLC - Reversed Phase:

  • Capcell PAK C18, IF, 1.8µm






Waters

UHPLC - Reversed Phase:

  • ACQUITY UPLC Oligonucleotide BEH C18, 1.7µm
  • ACQUITY Premier Oligonucleotide BEH C18, 1.7µm
  • ACQUITY Premier CSH, 1.7µm
  • XBridge Oligonucleotide BEH C18, 2.5µm
  • XTerra MS C18, 2.5µm

 

Sample preparation:

  • OASIS HLB SPE

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