Real-Time PCR for Gene Expression Quantification in Asthma

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 usi...

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Bibliographic Details
Published inMethods 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
LanguageEnglish
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
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Summary: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.
ISSN:1940-6029
DOI:10.1007/978-1-4939-3652-6_4