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Mol. Hum. Reprod. Advance Access originally published online on July 14, 2009
Molecular Human Reproduction 2009 15(11):757-761; doi:10.1093/molehr/gap058
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© The Author 2009. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

An RNA spiking method demonstrates that 18S rRNA is regulated by progesterone in the mouse uterus

R.G. Craythorn1,2, J.E. Girling2, M.P. Hedger1, P.A.W. Rogers2 and W.R. Winnall1,3

1Centre for Reproduction and Development, Monash Institute of Medical Research, Monash University, 27-31 Wright St, Clayton, VIC 3168, Australia 2Department of Obstetrics and Gynaecology and Monash Institute of Medical Research, Centre for Women's Health Research, Monash University, Clayton, VIC, Australia

3 Correspondence address. Tel: +61-3-9594-7426; Fax: +61-3-9594-7111; E-mail: wendy.winnall{at}med.monash.edu.au

Identifying suitable housekeeping genes for quantitative RT–PCR in the uterus is problematic, as this tissue undergoes significant structural and functional alterations during the oestrous cycle and pregnancy in response to circulating hormones. The suitability of 18S rRNA as a housekeeping gene in mouse uterus was investigated by introducing an ‘RNA spike’ standard into the reverse transcription reaction. 18S rRNA levels increased by Day 4 of pregnancy and after progesterone administration in ovariectomized mice. We conclude that 18S rRNA is not a suitable housekeeping gene for quantitative RT–PCR analysis in progesterone-responsive tissues, and the RNA spiking method provides a suitable alternative.

Key words: 18S rRNA/housekeeping gene/progesterone/quantitative real-time RT–PCR/uterus

Submitted on May 29, 2009; resubmitted on July 8, 2009; accepted on July 9, 2009.


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