Normalization of reverse transcription quantitative-PCR with housekeeping genes in rice

Biotechnol Lett. 2003 Nov;25(21):1869-72. doi: 10.1023/a:1026298032009.


Reverse transcription followed by real-time quantitative polymerase chain reaction (RT Q-PCR) is useful for the systematic measurement of plant physiological changes in gene expression. The validity of using 18S rRNA and three housekeeping genes, glyceraldehyde-3-phosphate dehydrogenase, actin, and tubulin, was tested as a reference of RT Q-PCR. Under various growth stages of etiolated seedlings, different cultivars, and various times after UV-irradiation treatment, expression level of 18S rRNA correlated with total RNA suggesting the uniformity of RT Q-PCR efficiencies among samples. Relative expressions of housekeeping genes varied among samples and independently of experimental conditions, up to two-fold, signifying generally constant fraction of mRNA in total RNA. Results indicate 18S rRNA was the most reliable reference gene for RT Q-PCR of total RNA.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Actins / genetics*
  • Actins / metabolism
  • Gene Expression Profiling / methods
  • Gene Expression Profiling / standards*
  • Gene Expression Regulation, Plant / genetics
  • Genetic Variation
  • Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / genetics*
  • Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / metabolism
  • Korea
  • Oryza / genetics*
  • Oryza / metabolism
  • Plant Proteins / genetics
  • Plant Proteins / metabolism
  • RNA, Ribosomal, 18S / genetics*
  • RNA, Ribosomal, 18S / metabolism
  • Reference Values
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Reverse Transcriptase Polymerase Chain Reaction / standards*
  • Tubulin / genetics*
  • Tubulin / metabolism


  • Actins
  • Plant Proteins
  • RNA, Ribosomal, 18S
  • Tubulin
  • Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating)