HPLC Method for Analysis of Oligonucleotides dA 5 mer on BIST A Column
Separation type: Bridge Ion Separation Technology, or BIST™ by SIELC Technologies
Synthesis of DNA Oligonucleotide (AAAAA)
The application of a DNA oligo with a sequence “AAAAA” can vary based on the context:
Using SIELC’s newly introduced BIST™ method, this oligonucleotide can be retained on a negatively-charged, cation-exchange BIST™ A column. There are two keys to this retention method: 1) a multi-charged, positive buffer, such as TMEDA formate, which acts as a bridge, linking the negatively charged dye to the negatively-charged column surface and 2) a mobile phase consisting mostly of organic solvent (such as MeCN) to minimize the formation of a solvation layer around the charged analytes. Using this new and unique analysis method, oligonucleotide can be separated, retained, and detected at 260 nm.
Methodenparameter
| Column | BIST A, 4.6 x 100 mm, 5 µm, 100 Å, surface coated |
|---|---|
| Mobile Phase | MeCN – 60% |
| Buffer | TMEDA Formate pH 4.0 – 20 mM |
| Flow Rate | 1.0 mL/min |
| Detection | UV 260 nm |
| Sample | 0.021 mg/ml in EtOH/H2O – 50/50% |
| Injection Volume | 1 µl |