High-speed preparative reversed-phase high-performance liquid chromatography of synthetic oligonucleotides.
A rapid method is described for the purification and analysis of synthetic oligonucleotides, based on reversed-phase high-performance liquid chromatography. Volatile buffers and a short column (40 mm X 4.6 mm) packed with Nucleosil 300-5 C4 were employed. Monitoring the column effluent with an UV detector provides an excellent means of controlling product quality. The method is suitable for the purification of crude synthesis products, as well as for desalting and removing gel contaminants from oligonucleotides eluted from polyacrylamide gels. The total time required per sample is less than 25 min.
Ashman
K
K
European Molecular Biology Laboratory, Heidelberg, F.R.G.
Bosserhoff
A
A
Frank
R
R
D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000032:analysis; D009841:Oligonucleotides / Q000302:isolation & purification*; D013056:Spectrophotometry, Ultraviolet
D009841:Oligonucleotides; D004247:DNA