Abstract
We have developed a method to isolate RNA in high yield from adult articular cartilage. Homogenization of the articular cartilage with a freezer mill, extraction with 4 M guanidinium isothiocyanate/acid-phenol, and ultracentrifugation in cesium trifluoroacetate was found to be an effective and practical method for isolating a high yield of intact RNA from adult canine articular cartilage. The total RNA was suitable for Northern blot analysis. The mRNA that could then be isolated by oligo-dT affinity chromatography was found to be a suitable substrate for in vitro translation, for making a cDNA library, and for PCR amplification.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Aggrecans
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Amino Acid Sequence
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Animals
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Autoradiography
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Base Sequence
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Blotting, Northern
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Cartilage, Articular / chemistry*
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Chromatography, Affinity
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Collagen / genetics*
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DNA Probes
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Dogs
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Electrophoresis, Polyacrylamide Gel
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Extracellular Matrix Proteins*
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Lectins, C-Type
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Molecular Sequence Data
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Nucleic Acid Hybridization
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Polymerase Chain Reaction
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Procollagen / genetics
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Protein Biosynthesis
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Proteoglycans / genetics
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RNA, Messenger / analysis
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RNA, Messenger / genetics
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RNA, Messenger / isolation & purification*
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Ultracentrifugation
Substances
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Aggrecans
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DNA Probes
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Extracellular Matrix Proteins
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Lectins, C-Type
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Procollagen
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Proteoglycans
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RNA, Messenger
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Collagen
Associated data
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GENBANK/M60299
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GENBANK/S37423
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GENBANK/S44552
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GENBANK/S51983