Evaluation of RNA from human trabecular bone and identification of stable reference genes.

Simona Cepollaro, Elena Della Bella, D. De Biase, M. Visani, Milena Fini

Research output: Contribution to journalArticlepeer-review

Abstract

The isolation of good quality RNA from tissues is an essential prerequisite for gene expression analysis to study pathophysiological processes. This study evaluated the RNA isolated from human trabecular bone and defined a set of stable reference genes. After pulverization, RNA was extracted with a phenol/chloroform method and then purified using silica columns. The A260/280 ratio, A260/230 ratio, RIN, and ribosomal ratio were measured to evaluate RNA quality and integrity. Moreover, the expression of six candidates was analyzed by qPCR and different algorithms were applied to assess reference gene stability. A good purity and quality of RNA was achieved according to A260/280 and A260/230 ratios, and RIN values. TBP, YWHAZ, and PGK1 were the most stable reference genes that should be used for gene expression analysis. In summary, the method proposed is suitable for gene expression evaluation in human bone and a set of reliable reference genes has been identified.
Original languageEnglish
Pages (from-to)4401-4407
Number of pages7
JournalJournal of Cellular Physiology
Volume233
Issue number6
DOIs
Publication statusPublished - Jun 2018

Keywords

  • RNA isolation
  • RNA quality
  • geNorm
  • gene expression
  • human bone
  • reference gene stability

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