Real-Time PCR for Gene Expression Quantification in Asthma

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1434; p. 45
• Main Authors: Segundo-Val, Ignacio San, García-Solaesa, Virginia, García-Sánchez, Asunción
• Format: Journal Article
• Language: English
• Published: United States 01.01.2016
• Subjects: Asthma - genetics; Fluorescent Dyes - chemistry; Gene Expression Profiling - methods; Gene Expression Regulation; Genetic Predisposition to Disease - genetics; Humans; Organic Chemicals - chemistry; Real-Time Polymerase Chain Reaction - methods; RNA, Messenger - analysis; RNA, Messenger - chemistry; Expression analysis; cDNA; mRNA; qPCR; RT-qPCR (reverse transcription-qPCR); Asthma
• DOI: 10.1007/978-1-4939-3652-6_4

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.