Selection of suitable reference genes for normalization of genes of interest in canine soft tissue sarcomas using quantitative real-time polymerase chain reaction.

Autor: Zornhagen KW; Department of Veterinary Clinical and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark.; Department of Clinical Physiology, Nuclear Medicine & PET, Copenhagen University Hospital and Cluster for Molecular Imaging, Faculty of Health Sciences, University of Copenhagen, Copenhagen N, Denmark., Kristensen AT; Department of Veterinary Clinical and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark., Hansen AE; Department of Clinical Physiology, Nuclear Medicine & PET, Copenhagen University Hospital and Cluster for Molecular Imaging, Faculty of Health Sciences, University of Copenhagen, Copenhagen N, Denmark.; Department of Micro- and Nanotechnology, Technical University of Denmark, Kgs. Lyngby, Denmark., Oxboel J; Department of Clinical Physiology, Nuclear Medicine & PET, Copenhagen University Hospital and Cluster for Molecular Imaging, Faculty of Health Sciences, University of Copenhagen, Copenhagen N, Denmark., Kjaer A; Department of Clinical Physiology, Nuclear Medicine & PET, Copenhagen University Hospital and Cluster for Molecular Imaging, Faculty of Health Sciences, University of Copenhagen, Copenhagen N, Denmark.
Jazyk: angličtina
Zdroj: Veterinary and comparative oncology [Vet Comp Oncol] 2015 Dec; Vol. 13 (4), pp. 485-93. Date of Electronic Publication: 2014 Jul 04.
DOI: 10.1111/vco.12108
Abstrakt: Quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) is a sensitive technique for quantifying gene expression. Stably expressed reference genes are necessary for normalization of RT-qPCR data. Only a few articles have been published on reference genes in canine tumours. The objective of this study was to demonstrate how to identify suitable reference genes for normalization of genes of interest in canine soft tissue sarcomas using RT-qPCR. Primer pairs for 17 potential reference genes were designed and tested in archival tumour biopsies from six dogs. The geNorm algorithm was used to analyse the most suitable reference genes. Eight potential reference genes were excluded from this final analysis because of their dissociation curves. β-Glucuronidase (GUSB) and proteasome subunit, beta type, 6 (PSMB6) were most stably expressed with an M value of 0.154 and a CV of 0.053 describing their average stability. We suggest that choice of reference genes should be based on specific testing in every new experimental set-up.
(© 2014 John Wiley & Sons Ltd.)
Databáze: MEDLINE
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