Extraction and isolation of mRNA from adult articular cartilage

Mark E. Adams; Ding Qiu Huang; Lena Yue Yao; Linda J. Sandell

doi:10.1016/0003-2697(92)90211-O

Extraction and isolation of mRNA from adult articular cartilage

Mark E. Adams, Ding Qiu Huang, Lena Yue Yao, Linda J. Sandell

Department of Orthopedic Surgery

Research output: Contribution to journal › Article › peer-review

50 Scopus citations

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.

Original language	English
Pages (from-to)	89-95
Number of pages	7
Journal	Analytical Biochemistry
Volume	202
Issue number	1
DOIs	https://doi.org/10.1016/0003-2697(92)90211-O
State	Published - Apr 1992

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title = "Extraction and isolation of mRNA from adult articular cartilage",

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.",

author = "Adams, {Mark E.} and {Qiu Huang}, Ding and {Yue Yao}, Lena and Sandell, {Linda J.}",

note = "Funding Information: This work was supported by Roussel Canada, Inc and NIH R0136994 (L.J.S.). We are very thankful to Dr. Dick HeinegHrd for encouragement and helpful suggestions, to Drs. John Matyas and David Hart critically reviewing the manuscript, and to Melanie Skromeda Xuefen Fan for excellent technical assistance.",

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doi = "10.1016/0003-2697(92)90211-O",

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AU - Adams, Mark E.

AU - Qiu Huang, Ding

AU - Yue Yao, Lena

AU - Sandell, Linda J.

N1 - Funding Information: This work was supported by Roussel Canada, Inc and NIH R0136994 (L.J.S.). We are very thankful to Dr. Dick HeinegHrd for encouragement and helpful suggestions, to Drs. John Matyas and David Hart critically reviewing the manuscript, and to Melanie Skromeda Xuefen Fan for excellent technical assistance.

PY - 1992/4

Y1 - 1992/4

N2 - 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.

AB - 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.

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Extraction and isolation of mRNA from adult articular cartilage

Abstract

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