Reference gene selection for quantitative real-time polymerase chain reaction in Populus

Anal Biochem. 2011 Jan 15;408(2):337-9. doi: 10.1016/j.ab.2010.08.044. Epub 2010 Sep 19.

Abstract

Accurate quantification of gene expression with quantitative real-time polymerase chain reaction (qRT-PCR) relies on the choice of an appropriate reference gene. In this study, nine candidate reference genes were selected to study the expression stability for qRT-PCR normalization in adventitious rooting of Populus hardwood cuttings. geNorm, NormFinder, and BestKeeper analysis revealed that actin isoform B (ACT) was the most unstable gene across developmental stages, whereas elongation factor 1 alpha (EF1a) and 18S recombinant RNA (18S) emerged as the most appropriate reference genes for qRT-PCR analysis in this complex developmental process.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / genetics
  • Actins / standards
  • Base Sequence
  • Genes, Plant
  • Peptide Elongation Factor 1 / genetics
  • Peptide Elongation Factor 1 / standards
  • Populus / genetics*
  • RNA, Ribosomal, 18S / genetics
  • RNA, Ribosomal, 18S / standards
  • Reference Standards
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Reverse Transcriptase Polymerase Chain Reaction / standards

Substances

  • Actins
  • Peptide Elongation Factor 1
  • RNA, Ribosomal, 18S