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Reference loci for RT-qPCR analysis of differentiating human embryonic stem cells.

Authors :
Vossaert, Liesbeth
O'Leary, Thomas
Van Neste, Christophe
Heindryckx, Björn
Vandesompele, Jo
De Sutter, Petra
Deforce, Dieter
Source :
BMC Molecular Biology; 2013, Vol. 14 Issue 1, p1-7, 7p, 1 Black and White Photograph, 2 Charts, 1 Graph
Publication Year :
2013

Abstract

Background: Selecting stably expressed reference genes is essential for proper reverse transcription quantitative polymerase chain reaction gene expression analysis. However, this choice is not always straightforward. In the case of differentiating human embryonic stem (hES) cells, differentiation itself introduces changes whereby reference gene stability may be influenced. Results: In this study, we evaluated the stability of various references during retinoic acid-induced (2 microM) differentiation of hES cells. Out of 12 candidate references, beta-2-microglobulin, ribosomal protein L13A and Alu repeats are found to be the most stable for this experimental set-up. Conclusions: Our results show that some of the commonly used reference genes are actually not amongst the most stable loci during hES cell differentiation promoted by retinoic acid. Moreover, a novel normalization strategy based on expressed Alu repeats is validated for use in hES cell experiments. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
14712199
Volume :
14
Issue :
1
Database :
Complementary Index
Journal :
BMC Molecular Biology
Publication Type :
Academic Journal
Accession number :
91262077
Full Text :
https://doi.org/10.1186/1471-2199-14-21