RT-PCR for the pseudogene-free amplification of the glyceraldehyde-3-phosphate dehydrogenase gene (gapd)

Harper, Lucy V., Hilton, Anthony C. and Jones, Alan F. (2003). RT-PCR for the pseudogene-free amplification of the glyceraldehyde-3-phosphate dehydrogenase gene (gapd). Molecular and Cellular Probes, 17 (5), pp. 261-265.

Abstract

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is an enzyme which catalyses the conversion of glyceraldehyde-3-phosphate to 1,3 diphosphoglycerate. It is considered to be constitutively expressed in all cells, and as such the gene for GAPDH (gapd) is commonly used as a benchmark reference in expression studies. However, previous investigations have demonstrated that gapd may show altered gene expression in a number of disease states and under certain experimental conditions, suggesting that results of experiments using gapd as a control should be interpreted with caution. Furthermore, consideration must be given to the potential co-amplification of pseudogenes of gapd during RT-PCR. Here, we describe a method to avoid the amplification of contaminating pseudogenes through the design of primers that bind only to genuine gapd mRNA transcript. © 2003 Elsevier Ltd. All rights reserved.

Publication DOI: https://doi.org/10.1016/S0890-8508(03)00063-X
Divisions: Life & Health Sciences > Biosciences
Life & Health Sciences
Uncontrolled Keywords: gene expression,glyceraldehyde-3-phosphate dehydrogenase,pseudogene,quantitative reverse transcription polymerase chain reaction,Cell Biology,Molecular Biology
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Related URLs: http://www.scop ... tnerID=8YFLogxK (Scopus URL)
Published Date: 2003-10
Authors: Harper, Lucy V.
Hilton, Anthony C. ( 0000-0001-8025-5270)
Jones, Alan F.

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