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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.
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
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
- Gl Sciences InertSustain Amide Analysis of Oligonucleotides
- GL Sciences InertsSustain AX-C18 Analysi of Oligonucleotides
- GL Sciences InertSustain AX-C18 Analysis of Oligonucleotides Metal Free Hardware
- GL Sciences InertSustainSwift C18 Analysis of Oligonucleotides
- GL Sciences ProteoSil 200-C18 Analysis of Oligonucleotides
- GL Sciences MonoClad C18-HS Analysis of DNA Oligomers
- GL Sciences MonoClad C18-HS Analysis of Ribonucleic Acids (ssRNA)
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
- Imtakt Intrada Organic Acid Deoxyoligonucleotides
- Imtakt Intrada Organic Acid Oligonucleotides (Oligothymidylic Acid)
- Imtakt Intrada Organic Acid Oligonucleotides (Oligothymidylic Acid)
- Imtakt Intrada Organic Acid Oligonucleotides (Oligothymidylic Acid)
- Imtakt Presto FF-C18 Deoxyoligonucleotides
- Imtakt Presto FF-C18 Oligonucleotides (Oligothymidylic Acid)
- Imtakt Scherzo SW-C18 Oligonucleotides
- Imtakt Unison UK-C18 Deoxyoligonucleotides
- Imtakt Unison US-C18 Oligonucleotides
Macherey-Nagel
HPLC - IEX:
- Nucleogel SAX
HPLC - HILIC:
- Nucleodur HILIC, 5µm
HPLC - Reversed Phase:
- Nucleosil C18, 5µm
- Macherey-Nagel Nucleogel SAX Separation of Synthetic Oligonucleotides
- Macherey-Nagel Nucleosil C18 5-Dimethoxytritylated Oligonucleotides
- Macherey-Nagel Nucleosil C18 Oligothymidine Phosphates
- Macherey-Nagel Nucleosil C18 Separation of dA3TA8 and its Phosphorylated Derivatives
- Macherey-Nagel Nucleodur HILIC Cyclobutane Pyrimidine Dimer Nucleotides
Nacalai Tesque
HPLC - Reversed Phase:
- Cosmosil C18-EB, 3µm
- Cosmocore PBr, 2.6µm
- Cosmocore C18, 2.6µm
- Cosmocore Cholester, 2.6µm
- Naclai Tesque Cosmocore C18, Cholester, PBr Oligonucleotides (dA) 19S10
- Naclai Tesque Cosmocore C18, Cholester, PBr Oligonucleotides (dT) 19-S10
- Naclai Tesque Cosmocore C18, Cholester, PBr Oligonucleotides (dT) 19-S17,18
- Naclai Tesque Cosmocore PBr Oligonucleotides (dT) 19
- Naclai Tesque Cosmocore PBr Oligonucleotides (dA)
- Naclai Tesque Cosmocore PBr Oligonucleotides (dA) 19
- Naclai Tesque Cosmosil C18-EB Oligonucleotides
- Naclai Tesque Cosmosil C18-EB Oligonucleotides (dA) 20 Ladder
- Naclai Tesque Cosmosil C18-EB Oligonucleotides (dA) Ladder
- Naclai Tesque Cosmosil C18-EB Oligonucleotides (dT)
- Naclai Tesque Cosmosil C18-EB Oligonucleotides (dT)
- Naclai Tesque Cosmosil C18-EB Oligonucleotides
- Naclai Tesque Cosmosil C18-EB Oligonucleotides
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
- Sepax Bio-C18, Proteomix SAX Oligonucleotide Analysis
- Sepax Proteomix SAX Separation od Oligonucleotide and its degarded Fragments
- Sepax Proteimix SAX Separation of Oligonucleotides and its Impurities
- Sepax Proteomix SAX Short RNA Oligonucleotides
- Sepax Zenix SEC, Zenix-C, Proteomix RP, Proteomix SAX Oligonucleotide Separation
- Sepax Zenix-C SEC-100 Short Oligonucleotides Analysis
- Sepax Proteomix SAX Oligonucleotide Separation
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
- Shimadzu Oligonucleotide Therapeutics Solution Guide
- Shimadzu Shim-pack Scepter Claris C18 Simple Analysis of Impurities in Oligonucleotide Therapeutics
- Shimadzu Shim-pack Bio IEX Q-NP Analysis of Synthesized Oligonucleotides
- Shimadzu Shim-pack BIO IEX Q-NP Oligonucleotides Analysis
- Shimadzu Shim-pack Scepter Claris C18 Oligonucleotide Characterization for Quality Control
- Shimadzu Shim-pack Scepter Claris C18 An Oligonucleotide Impurity Analysis Workflow
- Shimadzu Shim-pack Scepter HD-C18 In-Pair Reversed Phase Separation and Identification of Oligonucleotides
- Shimadzu Shim-pack Scepter C18 Rapid UHPLC Analysis of DNA (12mer) and its Impurities
- Shimadzu Shim-pack Scepter Claris C18 Analysis of Synthetic DNA
- Shimadzu Shim-pack Scepter Claris C18 Qualitative Analysis of Synthetic DNA
Shodex
HPLC - HILIC:
- HILICpak VN-50 1D
- HILICpak VN-50 2D
- Shodex HILICpak VN-50 1D High Sensitive Analysis of Synthetic Oligo-DNA
- Shodex HILICpak VN-50 2D Analysis of 10 - 50mer Oligo DNAs
- Shodex HILICpak VN-50 2D LC/UC/MS Analysis of Oligo DNA
- Shodex HILICpak VN-50 2D Analysis of Oligo DNAs
- Shodex HILICpak VN-50 2D Analysis of Oligonucleotides and their Impurities
- Shodex HILICpak VN-50 2D Analysis of Oligonucleotides and their Impurities Truncated Oligonucleotides
- Shodex HILICpak VN-50 2D Analysis of Oligonucleotides and their Impurities Base Alteration
- Shodex HILICpak VN-50 2D Analysis of Phosphorotioated Oligo DNA
- Shodex HILICpak VN-50 2D Analysis of 2'-OMe and 2'-MOE Modified Phosphorotioated Oligo DNA
- Shodex HILICpak VN-50 2D Analysis of Various Oligonucleotides
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
- Waters ACQUITY Oligonucleotide BEH C18 Developing a Novel, Integrated LC-MS Workflow for High-Resolution Monitoring and Characterization of Oligonucleotides
- Waters ACQUITY Oligonucleotide BEH C18 Rapid Analysis of Synthetic mRNA Cap Structure Using Ion-Pairing RPLC
- Waters ACQUITY Oligonucleotide BEH C18 UPLC/MS Analysis of Interfering RNA Oligonucleotides
- Waters ACQUITY Oligonucleotide BEH C18 Real-Time Analysis of RNAI Duplexes
- Waters ACQUITY Oligonucleotide BEH C18 UPLC Synapt MS/MS Method for Structural Characterization of siRNA Oligonucleotides
- Waters ACQUITY Oligonucleotide BEH C18 Enhanced Resolution for Longer Oligonucleotide Analytes
- Waters ACQUITY Oligonucleotide BEH C18 Sensitive Bioanalysis of Antisense Oligonucleotides of Various Lengths and Modifications
- Waters Application Notebook Oligonucleotides
- Waters OASIS HLB Purification of DNA Oligonucleotides
- Waters OASIS HLsalting of DNA OligonucleotidesB De
- Waters XBridge Oligonucleotide BEH C18 HPLC and UPLC Columns for the Analysis of Oligonucleotides
- Waters XBridge Oligonucleotide BEH C18 Semi-preparative Scale Single-stranded RNA Purification
- Waters XTerra MS C18 Quality Control of Synthetic Oligonucleotides
- Waters XTerra MS C18 RP-HPLC Method for Purification of Dually-Labeled Synthetic Oligonucleotides
- Waters XTerra MS C18 LC/MS Analysis of Synthetic Oligonucleotides
- Waters XTerra MS C18 RP-HPLC Method for the Purification of Labeled Synthetic Oligonucleotides
- Waters XTerra MS C18 HPLC Purification of Long Synthetic Oligonucleotides
- Waters XTerra MS C18 Separation of RNA Oligonucleotides
- Waters XTerra Oligonucleotides Purification Applications Notebook
- Waters ACQUITY Oligonucleotide BEH C18 Real-Time Analysis of RNAI Duplexes
- Waters ACQUITY Premier CSH LC-MS Analysis of siRNA, Single Guide RNA and Impurities
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