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Effective isolation of high-quality total RNA from human adult articular cartilage
L A McKenna1, A Gehrsitz, S Söder
1Cartilage Research Group, Institute of Pathology, University of Erlangen-Nürnberg, Krankenhausstrasse 8-10, Erlangen, D-91054, Germany.
Abstract:
The isolation of large quantities of good-quality RNA from human articular cartilage has been a long-standing problem for researchers working with human articular cartilage. In this paper we report a protocol which we have developed based on the Qiagen RNeasy procedure to produce high yields of purified, DNA-free RNA from normal and osteosteoarthritic human articular cartilage. The average yield of RNA was 8.39 microg/g (n = 59) for normal and 6.69 microg/g (n = 58) for osteoarthritic cartilage (average ratio OD 260/280 = 1.8-1.9). Quantitative PCR, cDNA array technology, and Northern blot analysis were used to verify the quality of the RNA.