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Large‐scale purification of oligonucleotides by extraction and precipitation with butanole
Author(s) -
Wolfrum Christian,
Josten Andre,
Bauer Georg,
Götz Peter
Publication year - 2004
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20378
Subject(s) - oligonucleotide , yield (engineering) , extraction (chemistry) , chromatography , precipitation , nucleic acid , chemistry , dna , biochemistry , materials science , physics , meteorology , metallurgy
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 μmol 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. © 2004 Wiley Periodicals, Inc.