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Nucleic Acids Research, 1994, Vol. 22, No. 10 1880-1884
© 1994

METHODS

Hybridization-based affinity partitioning of nucleic acids using PEG-coupled oligonucleotides

Andres Jäschke+,, Jens P. Fürste1, Volker A. Erdmann1 and Dieter Cech*,

Institut für Organische und Bioorganische Chemie, Humboldt-Universität zu Berlin Hessische Strasse 1-2, 10099 Berlin 1Institut für Biochemie, Freie Universität Berlin Thielallee 63, 14195 Berlin, Germany

*To whom correspondence should be addressed

Received December 27, 1993. Revised March 18, 1994. Accepted March 18, 1994.

Polyethylene glycol (PEG)-coupled oligonucleotides are partitioned in an aqueous two-phase system PEG/dextran. The affinity of the oligonucleotide for the PEG-rich phase increases proportionally to the length of the coupled PEG polymer. After hybridization, the PEG-coupled oligonucleotide is able to force a complementary nucleic acid strand into the PEG-rich phase. This property can be used for the sequence-specific isolation of nucleic acids through hybridization-based affinity partitioning. The dependence of the partition coefficient in this system on various parameters is described. The application of this principle to multistage chromatographic separations is demonstrated.

+Present address: Massachusetts Institute of Technology, Department of Biology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA


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