TY  - JOUR
TI  - Selection of reference genes in mouse embryos and in differentiating human and mouse ES cells
AU  - Willems, Erik
AU  - Mateizel, Ileana
AU  - Kemp, Caroline
AU  - Cauffman, Greet
AU  - Sermon, Karen
AU  - Leyns, Luc
T2  - The International Journal of Developmental Biology
AB  - Embryonic Stem (ES) cells have the potential to form every cell of the body and thus are of great promise for tissue transplantation. One of the rising techniques that allows studying the differentiation state of ES cells is quantitative RT-PCR (qRT-PCR). When relative quantification by qRT-PCR is applied, accurate normalization is necessary, since differentiated embryonic stem cells and developing embryos contain heterogeneous cell populations. Corrections for variations in the qRT-PCR reaction are needed to allow comparisons between different samples. We applied the normalization tools geNorm and Normfinder to ten reference genes identifying the most stable ones for relative quantification of gene expression during differentiation of human ES cells, as well as in differentiated mouse ES cells and in the developing mouse embryo. For relative quantification by qRT-PCR in these systems, we advise to use normalization factors based on multiple stable reference genes. However, when the use of several reference genes would be unpractical, a single reference gene in each experimental setup could be sufficient. When looking for single stable reference genes, beta-actin works best in both mouse embryo and ES cell experiments and glyceraldehyde-3-phosphate-dehydrogenase can be applied in both mouse and human ES cell experiments.
PY  - 2006
DO  - 10.1387/ijdb.052130ew
VL  - 50
IS  - 7
SP  - 627
EP  - 635
J2  - Int. J. Dev. Biol.
LA  - en
SN  - 0214-6282
SN  - 1696-3547
UR  - https://ijdb.ehu.eus/article/052130ew
Y2  - 2024/11/05/17:17:22
ER  -