Improved and Reproducible Flow Cytometry Methodology for Nuclei Isolation from Single Root Meristem
Joint Authors
Carvalho, Carlos Roberto
Abreu, Isabella Santiago
Silva, Thaís Cristina Ribeiro
Source
Issue
Vol. 2010, Issue 2010 (31 Dec. 2010), pp.1-7, 7 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2010-06-22
Country of Publication
Egypt
No. of Pages
7
Main Subjects
Abstract EN
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.
American Psychological Association (APA)
Silva, Thaís Cristina Ribeiro& Abreu, Isabella Santiago& Carvalho, Carlos Roberto. 2010. Improved and Reproducible Flow Cytometry Methodology for Nuclei Isolation from Single Root Meristem. Journal of Botany،Vol. 2010, no. 2010, pp.1-7.
https://search.emarefa.net/detail/BIM-463279
Modern Language Association (MLA)
Silva, Thaís Cristina Ribeiro…[et al.]. Improved and Reproducible Flow Cytometry Methodology for Nuclei Isolation from Single Root Meristem. Journal of Botany No. 2010 (2010), pp.1-7.
https://search.emarefa.net/detail/BIM-463279
American Medical Association (AMA)
Silva, Thaís Cristina Ribeiro& Abreu, Isabella Santiago& Carvalho, Carlos Roberto. Improved and Reproducible Flow Cytometry Methodology for Nuclei Isolation from Single Root Meristem. Journal of Botany. 2010. Vol. 2010, no. 2010, pp.1-7.
https://search.emarefa.net/detail/BIM-463279
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-463279