Abstract
Analysis of gene expression using real-time reverse transcription polymerase chain reaction (RT-PCR) requires reference genes to normalize expression values between samples. We have developed a novel reference for real-time RT-PCR using an arbitrary primer to amplify a random set of genes. The arbitrary primer amplifies over 30 genes, whose cumulative expression as measured by real-time RT-PCR closely follows that of UBQ 11, an Arabidopsis thaliana gene that is used as a reference on microarrays. The expression of arbitrary genes is also compared with potato (Solanum tuberosum spp. tubersosum) housekeeping genes and was shown to be stable during Phytophthora infestans infection.
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Abbreviations
- DNA:
-
deoxyribonucleic acid
- MoMLV:
-
Moloney murine leukemia virus
- RNA:
-
ribonucleic acid
- RT-PCR:
-
reverse transcription polymerase chain reaction
- DDRT-PCR:
-
differential display reverse-transcription polymerase chain reaction
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Acknowledgments
The authors would like to thank Ian Macdonald who provided technical assistance for this study.
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Tai, H.H., Conn, G., Davidson, C. et al. Arbitrary Multi-gene Reference for Normalization of Real-Time PCR Gene Expression Data. Plant Mol Biol Rep 27, 315–320 (2009). https://doi.org/10.1007/s11105-009-0089-0
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DOI: https://doi.org/10.1007/s11105-009-0089-0