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Nucleic Acids Research, 1979, Vol. 6, No. 6 2289-2306
© 1979

Articles

Reverse-phase HPLC of DNA restriction fragments and ribooligonucleotides on uncoated Kel-F powder

D.A. Usher

Department of Chemistry, Cornell University Ithaca, NY 14853, USA

Received February 12, 1979. Uncoated Kel-F powder offers some unique features as a support for reverse-phase HPLC of oligonucleotides and DNA restriction fragments. Compounds are eluted from the column by a gradient of acetonitrile ( 0 to 18% v/v) in 0.1 M aqueous triethylammonium acetate. In contrast to RPC-5 chromatography, oligonucleotides are not eluted by aqueous salt solutions alone, and the separation of restriction fragments depends only on the chainlength. The packing material is cheap, easy to pack, chemically inerr, and does not bleed, so that separations are highly reproducible. The DNA loading capacity for Kel-F is presently inferior to RPC-5, but recovery of µg amounts of material is typically better than 50%.


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