Validation of Zebrafish (Danio rerio) Reference Genes for Quantitative Real time RT PCR Normalization
Abstract The normalization of quantitative real time RT PCR (qRT PCR) is important to obtain accurate gene expression data. The most common method for qRT PCR normalization is to use reference, or house keeping genes. However, there is emerging evidence that even reference genes can be regulated under different conditions. qRT PCR has only recently been used in terms of zebrafish gene expression studies and there is no validated set of reference genes. This study characterizes the expression of nine possible reference genes during zebrafish embryonic development and in a zebrafish tissue panel. All nine reference genes exhibited variable expression. The b actin, EF1a and Rpl13a genes comprise a validated reference gene panel for zebrafish developmental time course studies, and the EF1a, Rpl13a and 18S rRNA genes are more suitable as a reference gene panel for zebrafish tissue analysis. Importantly, the zebrafish GAPDH gene appears unsuitable as reference gene for both types of studies.
Acta biochimica et biophysica Sinica