Article (Scientific journals)
Isolation of Nucleoli from Ehrlich Ascites Tumor Cells and Dynamics of Nascent RNA within Isolated Nucleoli.
Thiry, Marc; Ploton, Dominique
2008In Methods in Molecular Biology, 463, p. 111-21
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Abstract :
[en] Here we describe a new, rapid method for isolating nucleoli from Ehrlich tumor cells that preserves their morphological integrity and high transcriptional activity. Until now, methods for isolation of nucleoli were generally assumed to empty one of their three main compartments, the fibrillar center, of its contents. This new method consists of sonicating cells in an isotonic medium containing MgSO(4), spermidine, and spermine, followed by separation of nucleoli through a Percoll density gradient. Using the nonisotopic approach of labelling with BrUTP, we have further investigated the dynamics of nascent ribosomal RNAs (rRNAs) within morphologically intact isolated nucleoli at the electron microscope level. We show that ribosomal transcripts are elongated in the cortex of the fibrillar center and then enter the surrounding dense fibrillar component.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Thiry, Marc  ;  Université de Liège - ULiège > Département des sciences de la vie > Biologie cellulaire
Ploton, Dominique
Language :
English
Title :
Isolation of Nucleoli from Ehrlich Ascites Tumor Cells and Dynamics of Nascent RNA within Isolated Nucleoli.
Publication date :
2008
Journal title :
Methods in Molecular Biology
ISSN :
1064-3745
eISSN :
1940-6029
Publisher :
Humana Press, Totowa, United States - New Jersey
Volume :
463
Pages :
111-21
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 30 November 2009

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