Measurement of Cardiac Gene Expression by Reverse Transcription Polymerase Chain Reaction (RT-PCR)
By: Nicola King3
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Two methods applicable to the measurement of cardiac gene expression by reverse transcription polymerase chain reaction (RT-PCR)
are presented. Each method describes a specific technique and includes information on how to optimize the experiments and
how to trouble-shoot any problems. The first method illustrates how conventional RT-PCR was used to detect expression of a
specific gene in rat heart. The second method explains how to investigate the amount of gene expressed under different conditions
utilizing the new technique of quantitative real-time RT-PCR. More specifically, this second method was used to investigate
the effect of a relevant cardiac insult, ischemia reperfusion, on gene expression, although the technique could be readily
adapted to other cardiac insults or to compare different hearts or treatments. The gene investigated in these examples was
SNAT3, which is responsible for the cotransport of sodium predominantly with glutamine in exchange for hydrogen ions. The results
suggested that a 20-fold increase in SNAT3 expression occurs during ischemia, which reduced to baseline levels on reperfusion.
Affiliation(s): (3) Bristol Heart Institute, Department Clinical Science Medicine at South Bristol, Faculty of Medicine and Dentistry, University of Bristol, Bristol, UK
Book Title: Cardiac Gene Expression: Methods and Protocols
Series: Methods in Molecular Biology | Volume: 366 | Pub. Date: Mar-19-2007 | Page Range: 109-119 | DOI: 10.1007/978-1-59745-030-0_6
Subject: Genetics/Genomics
Key Words: RT-PCR - quantitative real-time RT-PCR - LightCycler™ - cardiac gene expression - ischemia reperfusion - amino acid transport -
SNAT3
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