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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.
The differences between oligonucleotide, nucleotide and DNA/RNA analysis using HPLC lie in the target molecules and the analysis methods:
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
AMT: HALO 120Å OLIGO C18, 2.7µm, 50x2.1mm
Competitor: OLIGO 130Å C18, 1.7µm, 50x2.1mm
Mobile Phase A: 100mM TEAA, pH 8.5
Mobile Phase B: Acetonitrile
Gradient:
Time / min | %B |
0 | 5 |
10 | 11 |
11 | 11 |
11.5 | 0 |
16.5 | 0 |
Flow rate: 0.5 mL/min
Back Pressure: HALO 140 bar (Competitor: 255 bar)
Temperature: 60 °C
Injection: 1.0 µL, 10 µg auf Säule
Sample Solvent: 10mM Tris HCl/1mM EDTA pH 8.0
Detection: PDA, 254nm
Flow Cell: 1 µL
Data Rate: 100 Hz
Response Time: 0.025 sec
LC System: Shimdazu Nexera X2
HPLC - Reversed Phase:
HPLC - IEX:
HPLC - HILIC:
HPLC - Reversed Phase:
UHPLC - Reversed Phase:
UHPLC - Reversed Phase:
HPLC - IEX:
HPLC - SEC:
HPLC - Reversed Phase:
UHPLC - Reversed Phase:
HPLC - Reversed Phase:
HPLC - IEX:
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