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 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.
McKenna
L A
LA
Cartilage Research Group, Institute of Pathology, University of Erlangen-Nürnberg, Krankenhausstrasse 8-10, Erlangen, D-91054, Germany.
Gehrsitz
A
A
Söder
S
S
Eger
W
W
Kirchner
T
T
Aigner
T
T
D000328:Adult; D015152:Blotting, Northern; D002358:Cartilage, Articular / Q000378:metabolism*; D016022:Case-Control Studies; D018076:DNA, Complementary / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D005821:Genetic Techniques; D006801:Humans; D007717:Knee; D020411:Oligonucleotide Array Sequence Analysis; D010003:Osteoarthritis / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011509:Proteoglycans / Q000378:metabolism; D012313:RNA / Q000032:analysis* / Q000302:isolation & purification*; D013053:Spectrophotometry
D018076:DNA, Complementary; D011509:Proteoglycans; D012313:RNA