RNA extraction from cartilage.

Détails

ID Serval
serval:BIB_C325F87E0323
Type
Partie de livre
Sous-type
Chapitre: chapitre ou section
Collection
Publications
Titre
RNA extraction from cartilage.
Titre du livre
Cartilage and Osteoarthritis. Volume 1: Cellular and Molecular Tools.
Auteur(s)
Mallein-Gerin F., Gouttenoire J.
Editeur
Sabatini M, Pastoureau P and De Ceuninck F
ISSN
1543-1894 (Print)
Statut éditorial
Publié
Date de publication
2004
Volume
100
Pages
101-104
Edition
Humana Press, Totowa, NJ
Langue
anglais
Notes
Publication types: Journal Article Publication Status: ppublish
Résumé
The direct isolation of RNA from cartilage has often proved difficult owing to a number of factors. Cartilage has a low cell content and contains an extracellular matrix rich in proteoglycans, which copurify with the RNA as they are large and negatively charged macromolecules. In our laboratory, we are interested in searching for genes differentially expressed in chondrocytes in diverse in vivo situations, for instance during maturation of chondrocytes in the growth plate or during cartilage degeneration. We found that treatment by proteinase K in 1 M guanidinium isothiocyanate prior to cesium trifluoroacetate ultracentrifugation was crucial to increase the yield and purity of RNA extracted from cartilage matrix. This protocol indeed led to reproducible patterns of differential display reverse transcriptase-polymerase chain reaction (RT-PCR) and should be useful for identifying genes differentially expressed by chondrocytes in situ.
Mots-clé
Animals, Cartilage/chemistry, Genetic Techniques, Humans, RNA/isolation & purification
Pubmed
Création de la notice
01/10/2015 15:34
Dernière modification de la notice
21/08/2019 5:34
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