Oligonucleotide Purification
Synthetic oligonucleotides cleaved from a solid support may contain failure sequences and other synthesis-related impurities. Reversed-phase solid-phase extraction(SPE) provides a convenient approach to cleanup and on-column detritylation before preparative HPLC, HPLC analysis, or LC/MS analysis. FUJIFILM Wako offers Presep® DNA/RNA Type A columns for simplified oligonucleotide purification.
How DMT-On Oligonucleotide Purification Works
Principle of DMT-On Oligonucleotide Purification by SPE
In oligonucleotide synthesis, the DMT (4,4′-dimethoxytrityl) group at the 5′ terminus may either be removed at the end of synthesis or retained for subsequent purification. When the DMT group is retained, reversed-phase solid-phase extraction (SPE) can be used for cleanup of the crude oligonucleotide product. The hydrophobic DMT-containing oligonucleotide is retained on the reversed-phase sorbent, while failure sequences and other synthesis-related impurities are washed away. The DMT group is then cleaved on the column under acidic conditions, using an acid such as trichloroacetic acid, trifluoroacetic acid, or dichloroacetic acid, followed by elution of the target oligonucleotide. Solid-phase extraction is therefore widely used as a straightforward method for the preliminary purification of synthetic oligonucleotides.
Solid-Phase Extraction Workflow for Oligonucleotide Purification
For research use or further manufacturing use only. Not for use in diagnostic procedures.
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