Large-scale purification of oligonucleotides by extraction and precipitation with butanole.
Today the synthesis of oligonucleotides is a well-established process. Using automatic synthesizers even kilogram quantities can be produced in a few hours. However, the purification of the final product is still time-consuming and needs a complex apparatus. In this article, a simple and fast purification method for the large-scale syntheses of oligonucleotides is described. According to the method of Sawadago and van Dyke ([1991] Nucleic Acids Res 19:674-675) for small-scale oligonucleotide purification, oligonucleotides in mumol to mmol amounts were purified by liquid-liquid extraction using butanole as the extraction liquid. Choosing appropriate ratios of extraction liquid to oligonucleotide solution, simultaneous purification and precipitation could be achieved. It was found that the yield of the purified oligonucleotide was mainly affected by the temperature. Yield decreased with increasing temperature. The use of this improved extraction procedure allows the purification of gram to kilogram quantities of oligonucleotides in less than a day with simple equipment and high yield.
Wolfrum
Christian
C
identif GmbH, Ulrich-Schalk-Str. 3, D-91056 Erlangen, Germany. wolfrum@identif.de
Josten
Andre
A
Bauer
Georg
G
Götz
Peter
P
D000440:Butanols / Q000737:chemistry*; D011232:Chemical Precipitation; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000302:isolation & purification*
D000440:Butanols; D009841:Oligonucleotides