Abstract
The quantitative real-time PCR (qPCR) has become the reference technique for studying gene expression in recent years. The application of qPCR to the study of asthma provides very useful information regarding the gene expression mechanisms. The quantification of RNA from cDNA can be performed by using fluorescent dyes or specific sequence probes. Here, we describe the protocol to quantify gene expression levels using SYBR Green as fluorescent dye. The protocol starts with the RNA extraction, followed by reverse transcription to obtain cDNA, quantification and finally data analysis.
An erratum to this chapter can be found at http://dx.doi.org/10.1007/978-1-4939-3652-6_19
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Acknowledgments
This work was supported by grants of the Junta de Castilla y León ref. GRS1047/A/14, GRS1189/A/15, and BIO/SA73/15, and by the project “Efecto del Ácido Retinóico en la enfermedad alérgica. Estudio transcripcional y su traslación a la clínica,” PI13/00564, integrated into the “Plan Estatal de I+D+I 2013–2016” and cofunded by the “ISCIII-Subdirección General de Evaluación y Fomento de la investigación” and the European Regional Development Fund (FEDER).
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San Segundo-Val, I., García-Solaesa, V., García-Sánchez, A. (2016). Real-Time PCR for Gene Expression Quantification in Asthma. In: Isidoro García, M. (eds) Molecular Genetics of Asthma. Methods in Molecular Biology, vol 1434. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3652-6_4
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DOI: https://doi.org/10.1007/978-1-4939-3652-6_4
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Online ISBN: 978-1-4939-3652-6
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