详细描述:
Oligonucleotides (whether synthetic or natural) may be purified by a number of techniques, including polyacrylamide gel electrophoresis and high performance liquid chromatography (either by ion exchange or reverse-phase methods). Synthetic DNA can be conveniently de-salted and purified by reverse-phase low-pressure separation on BTI's SuperPure Plus (SP-2000) columns . BTI's SuperPure Columns are intended for reverse-phase purification of 5'-DMT oligonucleotides followed by on-column detritylation. The method relies on strong binding between the 5'-DMT protected product and a hydrophobic optimized polymer packing (polystyrene), thus allowing separation from truncated sequences which, due to a lack of DMT group, do not bind. Treatment with acid removes the DMT group from the product, the desired product (free from organic residues) is eluted with 20% acetonitrile. The method is rapid, efficient and permits many samples to be purified simultaneously. Crude DNA from a 200 nmol synthesis scale can be loaded on the SuperPure Plus. The purified yield will depend, in each case, on sequence length and the quality of the crude sample.