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Gene expression analysis of shoot apical meristem cell types.

Methods in molecular biology (Clifton, N.J.) (2013-01-10)
Ram Kishor Yadav, Stephen Snipes, Thomas Girke, G Venugopala Reddy
ABSTRACT

Shoot apical meristems (SAMs) of higher plants harbor a set of stem-cells and provide cells for the development of all the above-ground biomass of plants. Most of the important pattern formation events such as maintenance of stem-cell identity, specification and differentiation of leaf/flower primordia, and temporal control of the transition from vegetative to reproductive program are determined in SAMs. Genetic analysis has revealed molecular and hormonal pathways involved in stem-cell maintenance, organ differentiation, and flowering time. However, limited information is available as to how different pathways interact with each other to function as a network in specifying different cell types and their function. Deciphering gene networks that underlie cell fate transitions requires new approaches aimed at assaying genome-scale expression patterns of genes at a single cell-type resolution. Here we provide details of experimental methods involved in protoplasting of SAM cells, generating cell type-specific gene expression profiles, and analysis platforms for identifying and inferring gene networks.

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)