Sedimentation field flow fractionation of DNA's.

Abstract

Sedimentation field flow fractionation (SFFF) is a method for purifying and providing mass or size distribution information on samples containing particulates or soluble macromolecules. Since SFFF separations are based on simple physical phenomena related to first principles, molecular weight (or particle sizes) can be determined without calibration standards. SFFF is a gentle technique suited for fractionating biomolecules. Studies with the fragile lambda DNA (molecular weight, 33 X 10(6] and smaller supercoiled plasmids have shown that these materials are not altered during SFFF separation; molecular weights and conformation remain unchanged, and biological activity is not reduced. Recoveries of nucleic acids approach 100 percent. Typically, components with about 20 percent difference in mass can be separated essentially to baseline if required. Fractionation time is usually independent of molecular weight, and separations often can be carried out within an hour.