Biotechnology Letters

, Volume 25, Issue 21, pp 1869–1872

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

  • Bo-Ra Kim
  • Hee-Young Nam
  • Soo-Un Kim
  • Su-Il Kim
  • Yung-Jin Chang
Article

DOI: 10.1023/A:1026298032009

Cite this article as:
Kim, B., Nam, H., Kim, S. et al. Biotechnology Letters (2003) 25: 1869. doi:10.1023/A:1026298032009

Abstract

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.

housekeeping genesOryza sativareverse transcription quantitative-PCRrice

Copyright information

© Kluwer Academic Publishers 2003

Authors and Affiliations

  • Bo-Ra Kim
    • 1
  • Hee-Young Nam
    • 1
  • Soo-Un Kim
    • 1
    • 2
  • Su-Il Kim
    • 1
  • Yung-Jin Chang
    • 1
  1. 1.School of Agricultural BiotechnologySeoul National UniversitySeoulKorea
  2. 2.Plant Metabolism Research CenterKyung Hee UniversitySuwonKorea