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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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Zenix-C SEC-100 3µm 100Å 300x7.8mm HPLC-Column
SPX-233100-7830
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Zenix SEC-300 3µm 300Å 300x7.8mm HPLC-Column
SPX-213300-7830
€2,190.00
Zenix SEC-150 3µm 150Å 300x7.8mm HPLC-Column
SPX-213150-7830
€2,190.00
Sepax Bio-C18 5µm 300Å 250x4.6mm HPLC-Column
SPX-106185-4625
€647.00
Sepax GP-C18 1.8µm 120Å 50x2.1mm HPLC-Column
SPX-101181-2105
€656.00
Cosmocore® 2.6PBr Packed Column 2.1mmx100mm
NT-13695-91
€599.00
Cosmocore® 2.6Cholester Packed Column 2.1mmx100mm
NT-12861-31
€601.00
Cosmocore® 2.6C18 Packed Column 2.1mmx100mm
NT-12614-71
Cosmosil® 3C18-EB Packed Column 2.0mmx100mm
NT-09796-01
Intrada Organic Acid, 150 mm x 2.0 mm LxID, 3µm, Intact organic acid LC-MS analysis colµmn
IMT-WOA25F
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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

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Manufacturer overview

AMT

HPLC - Reversed Phase:

  • HALO Oligo, 2.7µm

GL Sciences

HPLC - Reversed Phase:

  • ProteoSil 200-C18, 3µm
  • MonoClad C18-HS
  • InertSustainSwift C18, 3µm

 

HPLC - Mixed-Mode:

  • InertSustain AX-C18, 3µm

 

UHPLC - HILIC:

  • InertSustain Amide, 1.9µm

Hamilton

HPLC - Reversed Phase:

  • PRP-C18, 5µm

Imtakt

HPLC - Mixed-Mode:

  • Intrada Organic Acid, 3µm
  • Scherzo SW-C18, 3µm

 

HPLC - Reversed Phase:

  • Unison UK-C18, 3µm
  • Unison US-C18, 5µm

 

UHPLC - Reversed Phase:

  • Presto FF-C18, 2µm

Macherey-Nagel

HPLC - IEX:

  • Nucleogel SAX

 

HPLC - HILIC:

  • Nucleodur HILIC, 5µm

 

HPLC - Reversed Phase:

  • Nucleosil C18, 5µm

Nacalai Tesque

HPLC - Reversed Phase:

  • Cosmosil C18-EB, 3µm
  • Cosmocore PBr, 2.6µm
  • Cosmocore C18, 2.6µm
  • Cosmocore Cholester, 2.6µm

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

Sepax

HPLC - IEX:

  • Proteomix SAX, 3µm
  • Proteomix SAX, 5µm

 

HPLC - SEC:

  • Zenix SEC-300, 3µm
  • Zenix SEC-150, 3µm
  • Zenix-C SEC-150, 3µm
  • Zenix-C SEC-100, 3µm

 

HPLC - Reversed Phase:

  • Bio-C18, 5µm

 

UHPLC - Reversed Phase:

  • GP-C18, 1.8µm

Shimadzu

UHPLC - Reversed Phase:

  • Shim-pack Scepter Claris C18, 1.9µm
  • Shim-pack Scepter C18, 1.9µm
  • Shim-pack Scepter HD-C18, 1.9µm

 

HPLC - Reversed Phase:

  • Shim-pack Scepter C18, 5µm

 

HPLC - IEX:

  • Shim-pack Bio IEX Q-NP, 5µm

Shodex

HPLC - HILIC:

  • HILICpak VN-50 1D
  • HILICpak VN-50 2D

Thermo

HPLC - Reversed Phase:

  • DNAPac RP, 4µm

Tosoh

HPLC - IEX:

  • TSKgel DEAE-3SW, 10µm

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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