Abstract
Real-time, reverse transcription quantitative PCR (RT-qPCR) is a highly sensitive and reproducible technology for the analysis of gene expression patterns. Its ability to detect minute quantities of nucleic acid from multifarious sources makes it an ideal technique for embryonic transcript quantification. However, complex cellular diversity and active transcriptome dynamics in early embryos necessitate particular caution to avoid erroneous results. This chapter is intended to outline basic methodology to design and execute RT-qPCR experiments in pre-implantation embryos.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Bustin SA (2002) Quantification of mRNA using real-time reverse transcription PCR (RT-PCR): trends and problems. J Mol Endocrinol 29:23–39
Ståhlberg A, Håkansson J, Xian X et al (2004) Properties of the reverse transcription reaction in mRNA quantification. Clin Chem 50(3):509
Vandesompele J, De Preter K, Pattyn F et al (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 3(7):RESEARCH0034
Derveaux S, Vandesompele J, Hellemans J (2010) How to do successful gene expression analysis using real-time PCR. Methods 50(4):227–230
Bower NI, Moser RJ, Hill JR et al (2007) Universal reference method for real-time PCR gene expression analysis of preimplantation embryos. Biotechniques 42:199–206
Khan DK, Dube D, Gall L et al (2012) Expression of pluripotency master regulators during two key developmental transitions: EGA and early lineage specification in the bovine embryo. PLoS One 7(3):e34110
Yuan JS, Reed A, Chen F et al (2006) Statistical analysis of real-time PCR data. BMC Bioinformatics 7:85
Hellemans J, Mortier G, De Paepe A et al (2007) qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data. Genome Biol 8(2):19
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Peynot, N., Duranthon, V., Khan, D.R. (2015). Gene Expression Analysis in Early Embryos Through Reverse Transcription Quantitative PCR (RT-qPCR). In: Beaujean, N., Jammes, H., Jouneau, A. (eds) Nuclear Reprogramming. Methods in Molecular Biology, vol 1222. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1594-1_14
Download citation
DOI: https://doi.org/10.1007/978-1-4939-1594-1_14
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-1593-4
Online ISBN: 978-1-4939-1594-1
eBook Packages: Springer Protocols