Buoyant density centrifugation

Buoyant density centrifugation (also isopycnic centrifugation or equilibrium density-gradient centrifugation) uses the concept of buoyancy to separate molecules in solution by their differences in density.

Electron microscope image of canine parvovirus isolated using buoyant density centrifugation

Implementation

Historically a caesium chloride (CsCl) solution was often used, but more commonly used density gradients are sucrose or Percoll. The sample is put on top of the solution, and then the tube is spun at a very high speed for an extended time, at times lasting days. The CsCl molecules become densely packed toward the bottom, so even layers of different densities form. Since the original solution was approximately the same density, they go to a level where their density and the CsCl density are the same, to which they form a sharp, distinctive band.

Caesium chloride (CsCl) solution and two morphological types of rotavirus. Following centrifugation at 100g a density gradient forms in the CsCl solution and the virus particle separate according to their densities.

Isotope separation

This method very sharply separates molecules, and is so sharp that it can even separate different molecular isotopes from one another.

DNA separation

Buoyant density of majority of DNA is 1.7g/cm3 which is equal to density of 6M CsCl solution. Buoyant density of DNA changes with its GC content. The term "satellite DNA" refers to small bands of repetitive DNA sequences with distinct base composition floating above (A+T rich) or below (G+C rich) the main component DNA.

See also

References

    Further reading
    • Schildkraut, Carl L.; Marmur, Julius; Doty, Paul (1962). "Determination of the base composition of deoxyribonucleic acid from its buoyant density in CsCl". Journal of Molecular Biology. 4 (6): 430–443. doi:10.1016/S0022-2836(62)80100-4. ISSN 0022-2836.
    • James Greene (25 June 1998). Recombinant DNA Principles and Methodologies. CRC Press. pp. 278–. ISBN 978-0-8247-9989-2.


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