Selection of suitable reference genes for normalization of genes of interest in canine soft tissue sarcomas using quantitative real-time polymerase chain reaction
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Selection of suitable reference genes for normalization of genes of interest in canine soft tissue sarcomas using quantitative real-time polymerase chain reaction. / Zornhagen, Kamilla Westarp; Kristensen, Annemarie Thuri; Hansen, Anders Elias; Oxbøl, Jytte; Kjær, Andreas.
In: Veterinary and Comparative Oncology, Vol. 13, No. 4, 2015, p. 485–493.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Selection of suitable reference genes for normalization of genes of interest in canine soft tissue sarcomas using quantitative real-time polymerase chain reaction
AU - Zornhagen, Kamilla Westarp
AU - Kristensen, Annemarie Thuri
AU - Hansen, Anders Elias
AU - Oxbøl, Jytte
AU - Kjær, Andreas
N1 - © 2014 John Wiley & Sons Ltd.
PY - 2015
Y1 - 2015
N2 - 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.
AB - 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.
U2 - 10.1111/vco.12108
DO - 10.1111/vco.12108
M3 - Journal article
C2 - 24995963
VL - 13
SP - 485
EP - 493
JO - Veterinary and Comparative Oncology
JF - Veterinary and Comparative Oncology
SN - 1476-5810
IS - 4
ER -
ID: 131363700