Format

Send to

Choose Destination
Anal Biochem. 2017 Feb 1;518:134-138. doi: 10.1016/j.ab.2016.11.018. Epub 2016 Nov 29.

Extraction of high-quality RNA from human articular cartilage.

Author information

1
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.
2
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.

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.

PMID:
27913164
PMCID:
PMC5215757
DOI:
10.1016/j.ab.2016.11.018
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Elsevier Science Icon for PubMed Central
Loading ...
Support Center