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

Lucy V. Harper*, Anthony C. Hilton, Alan F. Jones

*Corresponding author for this work

Research output: Contribution to journalArticle

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.

Original languageEnglish
Pages (from-to)261-265
Number of pages5
JournalMolecular and Cellular Probes
Volume17
Issue number5
DOIs
Publication statusPublished - Oct 2003

Keywords

  • gene expression
  • glyceraldehyde-3-phosphate dehydrogenase
  • pseudogene
  • quantitative reverse transcription polymerase chain reaction

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