Phytoplasma pp 381-393 | Cite as

Cesium Chloride-Bisbenzimide Gradients for Separation of Phytoplasma and Plant DNA

  • Lucy T. T. Tran-NguyenEmail author
  • Bernd Schneider
Part of the Methods in Molecular Biology book series (MIMB, volume 938)


CsCl–bisbezimide gradient centrifugation is a method to separate phytoplasma from host plant DNA. Bisbenzimide forms a complex with A + T-rich DNA thereby lowering its relative density. During centrifugation the A + T-rich phytoplasma DNA is spatially separated from the less A + T-rich host plant DNA. The difference in buoyant density between phytoplasma DNA and plant DNA varies according to the host–pathogen combination. The phytoplasma DNA forms a distinct band above the host plant DNA and can be collected. Depending on the phytoplasma titer and the scale of extraction, highly purified DNA is obtained in sufficient quantities for the construction of a genomic library, a sequencing project or hybridization studies.

Key words

Bisbenzimide Buoyant density gradient centrifugation Cesium chloride DNA extraction Hoechst 33258 



The authors would like to thank Erich Seemüller (Julius Kühn-Institute, Federal Research Centre for Cultivated Plants, Institute for Plant Protection in Fruit Crops and Viticulture) for critically reading the chapter.


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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  1. 1.Department of ResourcesPlant IndustriesDarwinAustralia
  2. 2.Julius Kühn-Institute (JKI), Federal Research Centre for Cultivated Plants, Institute for Plant Protection in Fruit Crops and ViticultureDossenheimGermany

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