GAPDH Pseudogenes and the Quantification of Feline Genomic DNA Equivalents
Joint Authors
Helfer-Hungerbuehler, A. Katrin
Hofmann-Lehmann, Regina
Widmer, Stefan
Source
Molecular Biology International
Issue
Vol. 2013, Issue 2013 (31 Dec. 2013), pp.1-7, 7 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2013-04-28
Country of Publication
Egypt
No. of Pages
7
Main Subjects
Natural & Life Sciences (Multidisciplinary)
Biology
Abstract EN
Quantitative real-time PCR (qPCR) is broadly used to detect and quantify nucleic acid targets.
In order to determine cell copy number and genome equivalents, a suitable reference gene that is present in a defined number in the genome is needed, preferably as a single copy gene.
For most organisms, a variable number of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) pseudogenes have been reported.
However, it has been suggested that a single-copy of the GAPDH pseudogene is present in the feline genome and that a GAPDH assay can therefore be used to quantify feline genomic DNA (gDNA).
The aim of this study was to determine whether one or more GAPDH pseudogenes are present in the feline genome and to provide a suitable alternative qPCR system for the quantification of feline cell copy number and genome equivalents.
Bioinformatics and sequencing results revealed that not just one but several closely related GAPDH-like sequences were present in the cat genome.
We thus identified, developed, optimized, and validated an alternative reference gene assay using feline albumin (fALB).
Our data emphasize the need for an alternative reference gene, apart from the GAPDH pseudogene, for the normalization of gDNA levels.
We recommend using the fALB qPCR assay for future studies.
American Psychological Association (APA)
Helfer-Hungerbuehler, A. Katrin& Widmer, Stefan& Hofmann-Lehmann, Regina. 2013. GAPDH Pseudogenes and the Quantification of Feline Genomic DNA Equivalents. Molecular Biology International،Vol. 2013, no. 2013, pp.1-7.
https://search.emarefa.net/detail/BIM-483072
Modern Language Association (MLA)
Helfer-Hungerbuehler, A. Katrin…[et al.]. GAPDH Pseudogenes and the Quantification of Feline Genomic DNA Equivalents. Molecular Biology International No. 2013 (2013), pp.1-7.
https://search.emarefa.net/detail/BIM-483072
American Medical Association (AMA)
Helfer-Hungerbuehler, A. Katrin& Widmer, Stefan& Hofmann-Lehmann, Regina. GAPDH Pseudogenes and the Quantification of Feline Genomic DNA Equivalents. Molecular Biology International. 2013. Vol. 2013, no. 2013, pp.1-7.
https://search.emarefa.net/detail/BIM-483072
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-483072