Use este identificador para citar ou linkar para este item: https://locus.ufv.br//handle/123456789/17251
Tipo: Artigo
Título: Improved and reproducible flow cytometry methodology for nuclei isolation from single root meristem
Autor(es): Silva, Thaís Cristina Ribeiro
Abreu, Isabella Santiago
Carvalho, Carlos Roberto
Abstract: Root meristems have increasingly been target of cell cycle studies by flow cytometric DNA content quantification. Moreover, roots can be an alternative source of nuclear suspension when leaves become unfeasible and for chromosome analysis and sorting. In the present paper, a protocol for intact nuclei isolation from a single root meristem was developed. This proceeding was based on excision of the meristematic region using a prototypical slide, followed by short enzymatic digestion and mechanical isolation of nuclei during homogenization with a hand mixer. Such parameters were optimized for reaching better results. Satisfactory nuclei amounts were extracted and analyzed by flow cytometry, producing histograms with reduced background noise and CVs between 3.2 and 4.1%. This improved and reproducible technique was shown to be rapid, inexpensive, and simple for nuclear extraction from a single root tip, and can be adapted for other plants and purposes.
Palavras-chave: cell
Nuclei isolation
Editor: Journal of Botany
Tipo de Acesso: Open Access
URI: http://dx.doi.org/10.1155/2010/320609
http://www.locus.ufv.br/handle/123456789/17251
Data do documento: 6-Mai-2010
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