Sedimentation field flow fractionation of DNA's
Schallinger, L.E.; Yau, W.W.; Kirkland, J.J.
Science 225(4660): 434-437
1984
ISSN/ISBN: 0036-8075 PMID: 6377495 Document Number: 223995
Sedimentation field flow fractionation (SFFF) is a method for purifying and providing mass or size distribution information on samples containing particulates or soluble macromolecues. Since SFFF separations are based on simple physical phenomena related to first principles, MW (or particle sizes) can be determined without calibration standards. SFFF is a gentle technique suited for fractionating biomolecules. Studies with the fragile .lambda. DNA (MW, 33 .times. 106) and smaller supercoiled plasmids showed that these materials are not altered during SFFF separation; MW and conformation remain unchanged, and biological activity is not reduced. Recoveries of nucleic acids approach 100%. Typically, components with about 20% difference in mass can be separated essentially to baseline if required. Fractionation time is usually independent of MW, and separations often can be carried out within an hour.