Effective isolation of high-quality total RNA from human adult articular cartilage

The isolation of large quantities of good-quality RNA from human articular cartilage has been a long-standing problem for researchers working with hum an articular cartilage. In this paper we report a protocol which we have de veloped based on the Qiagen RNeasy procedure to produce high yields of puri fied, RNA-free RNA from normal and osteosteoarthritic human articular carti lage. The average yield of RNA was 8.39 mug/g (n = 59) for normal and 6.69 mug/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. (C) 2000 Academic Press.