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.
Publication types
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Comparative Study
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Evaluation Study
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Research Support, Non-U.S. Gov't
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Validation Study
MeSH terms
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Actins / genetics*
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Actins / metabolism
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Gene Expression Profiling / methods
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Gene Expression Profiling / standards*
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Gene Expression Regulation, Plant / genetics
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Genetic Variation
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Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / genetics*
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Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / metabolism
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Korea
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Oryza / genetics*
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Oryza / metabolism
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Plant Proteins / genetics
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Plant Proteins / metabolism
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RNA, Ribosomal, 18S / genetics*
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RNA, Ribosomal, 18S / metabolism
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Reference Values
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Reverse Transcriptase Polymerase Chain Reaction / methods*
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Reverse Transcriptase Polymerase Chain Reaction / standards*
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Tubulin / genetics*
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Tubulin / metabolism
Substances
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Actins
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Plant Proteins
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RNA, Ribosomal, 18S
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Tubulin
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Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating)