Abstract
Here we describe methods for producing nuclei from Arabidopsis suspension cultures or root tips of Arabidopsis, wheat, or pea. These methods could be adapted for other species and cell types. The resulting nuclei can be further purified for use in biochemical or proteomic studies, or can be used for microscopy. We also describe how the nuclei can be used to obtain a preparation of nucleoli.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Pendle AF, Clark GP, Boon R et al (2005) Proteomic analysis of the Arabidopsis nucleolus suggests novel nucleolar functions. Mol Biol Cell 16:260–269
McKeown P, Pendle AF, Shaw PJ (2008) Preparation of Arabidopsis nuclei and nucleoli. Methods Mol Biol 463:67–75
Pendle AF, Shaw PJ (2015) Immunolabelling and in situ labelling of isolated plant interphase nuclei. Methods Mol Biol. In press
Saxena PK, Fowke LC, King J (1985) An efficent procedure for the isolation of nuclei from plant protoplasts. Protoplasma 128:184–189
Jackson DA, Cook PR (1985) A general method for preparing chromatin containing intact DNA. EMBO J 4:913–918
Staudt T, Lang MC, Medda R et al (2007) 2,2’-Thiodiethanol: a new water soluble mounting medium for high resolution optical microscopy. Microsc Res Tech 70:1–9
Acknowledgements
This work was supported by grant BB/J004588/1 from BBSRC and the John Innes Foundation.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media New York
About this protocol
Cite this protocol
Pendle, A.F., Shaw, P.J. (2017). Isolation of Nuclei and Nucleoli. In: Taylor, N., Millar, A. (eds) Isolation of Plant Organelles and Structures. Methods in Molecular Biology, vol 1511. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6533-5_3
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6533-5_3
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6531-1
Online ISBN: 978-1-4939-6533-5
eBook Packages: Springer Protocols