Selection of Suitable Reference Genes for Quantitative Real-Time PCR Normalization in Human Stem Cell Research

Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 1119)


Quantitative real-time polymerase chain reaction (qRT-PCR) is a widely utilized method for evaluating the gene expressions in stem cell research. This method enables researchers to obtain fast and precise results, but the accuracy of the data depends on certain factors, such as those associated with biological sample preparation and PCR efficiency. In order to achieve accurate and reliable results, it is of utmost importance to designate the reference genes, the expressions of which are suitable to all kinds of experimental conditions. Hence it is vital to normalize the qRT-PCR data by using the reference genes. In recent years, it has been found that the expression levels of reference genes widely used in stem cell research present a substantial amount of variation and are not necessarily suitable for normalization. This chapter at hand stresses the significance of selecting suitable reference genes from the point view of human stem cell research.


Reference genes qRT-PCR normalization qRT-PCR Stem cells Human Housekeeping genes 


18S rRNA

18S Ribosomal RNA






3′ untranslated region of the genes


Actin beta


5′-Aminolevulinate synthase 1


Adipose stem cells




Bone marrow-derived mesenchymal stem cells


Calpain 10


Cancer susceptibility 3


Cilia and flagella associated protein 65


Cancer stem cells


Eukaryotic translation elongation factor 1 alpha 1


EP300 interacting inhibitor of differentiation 2


Embryonic stem cells


F-Box and leucine rich repeat protein 12


Fetal tissue-derived MSCs


Growth arrest and DNA damage inducible alpha


Glyceraldehyde-3-phosphate dehydrogenase


Gingival stem cells


Glucuronidase beta




Hydroxymethylbilane synthase


Hypoxanthine phosphoribosyltransferase 1


Hematopoietic stem cells


Importin 8


Induced pluripotent stem cells


Leukotriene B4 receptor 2


Marrow-isolated adult multilineage inducible


Mesenchymal stem cells


Nucleotide binding protein 1


Polymerase chain reaction


Phosphoglycerate kinase


Peptidylprolyl isomerase A


Pumilio RNA binding family member 1


Quantitative real-time PCR


Rabaptin, RAB GTPase binding effector protein 2


Ring finger protein 7


Ribosomal protein L13a


Ribosomal protein 19


Ribosomal protein lateral stalk subunit P0


Ribosomal protein S18


Succinate dehydrogenase complex flavoprotein subunit A


Solute carrier family 4 member 1 adaptor protein


Signal recognition particle 72


TATA-box binding protein


Transferrin receptor


TNF receptor superfamily member 13C




Vascular endothelial growth factor A-165


Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta


Zinc finger protein 324B


Competing Interests

Y.M.E. is the founder and director of Biovalda Health Technologies, Inc. (Ankara, Turkey). The authors declare no competing interests in relation to this article.


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© Springer Nature Switzerland AG 2018

Authors and Affiliations

  1. 1.Tissue Engineering, Biomaterials and Nanobiotechnology LaboratoryAnkara University Faculty of Science, and Ankara University Stem Cell InstituteAnkaraTurkey
  2. 2.Biovalda Health Technologies, Inc.AnkaraTurkey

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