Graciela Jannice
Normalization, which is typically accomplished by comparing the abundance of the gene of interest to that of an endogenous reference gene, is an essential step in gene expression analysis for obtaining useful data from reverse transcription quantitative PCR (RT qPCR) assays. It's not easy to find these reference genes, which should be stable when expressed in multiple tissue samples and under different experimental conditions. A set of genes has been identified and evaluated in this work that could serve as a reference for gene expression studies on water buffalo. The first step involves downloading a Bos taurusexpressed sequence tags database from the TIGR Gene Index and mining it with simple frequency algorithms to determine which tentative consensuses are more suitable for inclusion in a starter set of candidate reference genes because they are present in a remarkable number of different cDNA libraries. An RT qPCR analysis, in which the expression stability of these genes was evaluated on a panel of buffalo tissues and organs, was carried out in order to validate the potential of such candidates for their use as normalizers in buffalo gene expression analysis. According to our findings, gene expression levels in buffalo tissues and organs can be compared using normalizers based on ribosomal proteins L4 and L5 and Gek proteins.
KeywordsGene expression; Reference genes; Normalization; RT qPCR
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