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 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.
Department of Clinical Biochemistry, University Hospital of Salamanca, Salamanca, Spain. i.sansegundo.val@gmail.com.|Salamanca Institute for Biomedical Research (IBSAL), Salamanca, Spain. i.sansegundo.val@gmail.com.
Ignacio San
IS
Segundo-Val
Salamanca Institute for Biomedical Research (IBSAL), Salamanca, Spain.|Department of Clinical Genetics, University Hospital of Navarra, Pamplona, Navarra, Spain.
Virginia
V
García-Solaesa
Department of Biomedical and Diagnostic Sciences, University of Salamanca, Salamanca, Spain.
Asunción
A
García-Sánchez
D001249:Asthma / Q000235:genetics*; D052160:Benzothiazoles; D003959:Diamines; D005456:Fluorescent Dyes / Q000737:chemistry; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation; D020022:Genetic Predisposition to Disease / Q000235:genetics; D006801:Humans; D009930:Organic Chemicals / Q000737:chemistry; D011804:Quinolines; D012333:RNA, Messenger / Q000032:analysis / Q000737:chemistry; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
D052160:Benzothiazoles; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; D012333:RNA, Messenger; C098022:SYBR Green I
Asthma; Expression analysis; RT-qPCR (reverse transcription-qPCR); cDNA; mRNA; qPCR