Extraction of high-quality RNA from human articular cartilage

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• 作者：Heather K. Le Bleu^a ; ^b ; Fadia A. Kamal^a ; ^b ; Meghan Kelly^a ; ^b ; John P. Ketz^a ; ^b ; Michael J. Zuscik^a ; ^b ; Reyad A. Elbarbary^a ; ^b ; ^c ; ^d ; ^{reyad_elbarbary@urmc.rochester.edu}
• 刊名：Analytical Biochemistry
• 出版年：2017
• 出版时间：1 February 2017
• 年：2017
• 卷：518
• 期：Complete
• 页码：134-138
• 全文大小：1150 K
• 卷排序：518

文摘

Extracting high-quality RNA from articular cartilage is challenging due to low cellularity and high proteoglycan content. This problem hinders efficient application of RNA sequencing (RNA-seq) analysis in studying cartilage homeostasis. Here we developed a method that purifies high-quality RNA directly from cartilage. Our method optimized the collection and homogenization steps so as to minimize RNA degradation, and modified the conventional TRIzol protocol to enhance RNA purity. Cartilage RNA purified using our method has appropriate quality for RNA-seq experiments including an RNA integrity number of ∼8. Our method also proved efficient in extracting high-quality RNA from subchondral bone.