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Isolation of Nuclei Tagged in Specific Cell Types (INTACT) in Arabidopsis

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Flower Development

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2686))

Abstract

Many functionally distinct plant tissues have relatively low numbers of cells that are embedded within complex tissues. For example, the shoot apical meristem (SAM) consists of a small population of pluripotent stem cells surrounded by developing leaves and/or flowers at the growing tip of the plant. It is technically challenging to collect enough high-quality SAM samples for molecular analyses. Isolation of Nuclei Tagged in specific Cell Types (INTACT) is an easily reproducible method that allows the enrichment of biotin-tagged cell-type-specific nuclei from the total nuclei pool using biotin-streptavidin affinity purification. Here, we provide a detailed INTACT protocol for isolating nuclei from the Arabidopsis SAM. One can also adapt this protocol to isolate nuclei from other tissues and cell types for investigating tissue/cell-type-specific transcriptome and epigenome and their changes during developmental programs at a high spatiotemporal resolution. Furthermore, due to its low cost and simple procedures, INTACT can be conducted in any standard molecular laboratory.

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Author information

Authors and Affiliations

  1. Umeå Plant Science Centre, Department of Plant Physiology, Umeå University, Umeå, Sweden

    Ruben M. Benstein & Markus Schmid

  2. Center for Plant Molecular Biology (ZMBP), Department of General Genetics, Eberhard Karls University of Tübingen, Tübingen, Germany

    Yuan You

  3. Department of Plant Biology, Linnean Centre for Plant Biology, Swedish University of Agricultural Sciences, Uppsala, Sweden

    Markus Schmid

  4. Department of Molecular Life Sciences, Technical University of Munich, Freising, Germany

    Yuan You

Authors
  1. Ruben M. Benstein
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  2. Markus Schmid
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  3. Yuan You
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Corresponding author

Correspondence to Yuan You .

Editor information

Editors and Affiliations

  1. Centre for Research in Agricultural Genomics CSIC-IRTA-UAB-UB, Barcelona, Spain

    José Luis Riechmann

  2. Instituto de Biología Molecular y Celular de Plantas CSIC-UPV, Valencia, Spain

    Cristina Ferrándiz

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Benstein, R.M., Schmid, M., You, Y. (2023). Isolation of Nuclei Tagged in Specific Cell Types (INTACT) in Arabidopsis. In: Riechmann, J.L., Ferrándiz, C. (eds) Flower Development . Methods in Molecular Biology, vol 2686. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3299-4_16

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  • DOI: https://doi.org/10.1007/978-1-0716-3299-4_16

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3298-7

  • Online ISBN: 978-1-0716-3299-4

  • eBook Packages: Springer Protocols

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