Extraction of high-quality RNA from human articular cartilage

Heather K. Le Bleu, Fadia A. Kamal, Meghan Kelly, John P. Ketz, Michael J. Zuscik, Reyad A. Elbarbary

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)134-138
Number of pages5
JournalAnalytical Biochemistry
Volume518
DOIs
StatePublished - Feb 1 2017

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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