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  • Fluorescence activated cell sorting of shoot apical meristem cell types.

Fluorescence activated cell sorting of shoot apical meristem cell types.

Methods in molecular biology (Clifton, N.J.) (2014-01-08)
G Venugopala Reddy
ABSTRACT

Growing tips of plants harbor a set of stem cells in structures called shoot apical meristems (SAMs) which provide cells for development of aboveground biomass. Despite a periodic differentiation of stem cell progenitors into leaves, the stem cell pool remains constant over time. Genetic analysis has revealed molecular pathways involved in stem-cell specification, cell division patterns, and organ differentiation. Stem cells within SAMs are few in number, which imposes a limitation to the experimental approaches that can be used for deciphering the gene regulatory networks that underlie cell fate transitions. Here, I provide detailed experimental protocols for the protoplasting and subsequent purification through cell sorting of SAM cells, which allows genome-wide analyses of gene expression patterns at a single cell-type resolution.

MATERIALS
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Sigma-Aldrich
Hemicellulase from Aspergillus niger, powder, 0.3-3.0 unit/mg solid (using a β-galactose dehydrogenase system and locust bean gum as substrate)