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

Adv Exp Med Biol. 2018;1119:151-168. doi: 10.1007/5584_2018_277.

Abstract

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.

Keywords: Housekeeping genes; Human; Reference genes; Stem cells; qRT-PCR; qRT-PCR normalization.

MeSH terms

  • Gene Expression Profiling*
  • Gene Expression Regulation, Plant
  • Humans
  • Real-Time Polymerase Chain Reaction
  • Reference Standards
  • Stem Cell Research*