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

Ruben M Benstein, Markus Schmid, Yuan You

Methods in molecular biology (Clifton, N.J.) 2023

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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. © 2023. Springer Science+Business Media, LLC, part of Springer Nature.

Citation

Ruben M Benstein, Markus Schmid, Yuan You. Isolation of Nuclei Tagged in Specific Cell Types (INTACT) in Arabidopsis. Methods in molecular biology (Clifton, N.J.). 2023;2686:313-328