title: Extraction of high-quality RNA from human articular cartilage.
abstract: 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.
authors.affiliation: Center for Musculoskeletal Research, School of Medicine and Dentistry, University of Rochester, Rochester, NY 14642, USA; Department of Orthopedics and Rehabilitation, School of Medicine and Dentistry, University of Rochester, Rochester, NY 14642, USA.
authors.forename: Heather K
authors.initials: HK
authors.lastname: Le Bleu
authors.affiliation: Center for Musculoskeletal Research, School of Medicine and Dentistry, University of Rochester, Rochester, NY 14642, USA; Department of Orthopedics and Rehabilitation, School of Medicine and Dentistry, University of Rochester, Rochester, NY 14642, USA.
authors.forename: Fadia A
authors.initials: FA
authors.lastname: Kamal
authors.affiliation: Center for Musculoskeletal Research, School of Medicine and Dentistry, University of Rochester, Rochester, NY 14642, USA; Department of Orthopedics and Rehabilitation, School of Medicine and Dentistry, University of Rochester, Rochester, NY 14642, USA.
authors.forename: Meghan
authors.initials: M
authors.lastname: Kelly
authors.affiliation: Center for Musculoskeletal Research, School of Medicine and Dentistry, University of Rochester, Rochester, NY 14642, USA; Department of Orthopedics and Rehabilitation, School of Medicine and Dentistry, University of Rochester, Rochester, NY 14642, USA.
authors.forename: John P
authors.initials: JP
authors.lastname: Ketz
authors.affiliation: Center for Musculoskeletal Research, School of Medicine and Dentistry, University of Rochester, Rochester, NY 14642, USA; Department of Orthopedics and Rehabilitation, School of Medicine and Dentistry, University of Rochester, Rochester, NY 14642, USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Zuscik
authors.affiliation: Center for Musculoskeletal Research, School of Medicine and Dentistry, University of Rochester, Rochester, NY 14642, USA; Department of Orthopedics and Rehabilitation, School of Medicine and Dentistry, University of Rochester, Rochester, NY 14642, USA; Department of Biochemistry and Biophysics, School of Medicine and Dentistry, University of Rochester, Rochester, NY 14642, USA; Center for RNA Biology, University of Rochester, Rochester, NY 14642, USA. Electronic address: reyad_elbarbary@urmc.rochester.edu.
authors.forename: Reyad A
authors.initials: RA
authors.lastname: Elbarbary
mesh_terms: D002358:Cartilage, Articular / Q000737:chemistry*; D006801:Humans; D007719:Knee Joint / Q000737:chemistry*; D008297:Male; D012313:RNA / Q000737:chemistry* / Q000302:isolation & purification*
chemical_list: D012313:RNA