Abstract
BackgroundReference genes are usually stably expressed in various cells and tissues. However, it was reported that the expression of some reference genes may be distinct in different species. In this study, we intend to answer whether the expression of reported traditional reference genes changes or not in the polyploid fishResultsBy retrieving the mRNA sequencing data of three different ploidy fish from the NCBI SRA database, we selected 12 candidate reference genes, and examined their expression levels in the 10 tissues and in the four cell lines of three different ploidy fish by real-time PCR. Then, the expression profiles of these 12 candidate reference genes were systematically evaluated by using the software platforms: BestKeeper, NormFinder and geNorm.ConclusionThe 28S ribosomal protein S5 gene (RPS5) and the ribosomal protein S18 gene (RPS18) are the most suitable reference genes for the polyploid of Cyprinus carpio and Carassius auratus, demonstrated by both of the tissues and the cultured cells.
Highlights
Reference genes are usually stably expressed in various cells and tissues
A number of reference genes have been widely used in molecular biology research, such as the beta-actin (β-actin), beta-tubulin (β-tubulin), elongation factor 1-alpha (EF1-α), Glyceraldehyde-3phosphate dehydrogenase (GAPDH), ribosomal protein S18 (RPS18), etc. [3,4,5,6,7]
Based on the transcriptome data, we found that the expression of some traditional reference genes was unstable in polyploidy, such as the β-actin
Summary
Reference genes are usually stably expressed in various cells and tissues. it was reported that the expression of some reference genes may be distinct in different species. We intend to answer whether the expression of reported traditional reference genes changes or not in the polyploid fish. It was reported that the expression of some traditional reference genes could change in varying degrees in various species or under. Polyploids are the organisms containing three or more complete sets of chromosomes [17]. They are widespread in plants [18,19,20,21,22,23], and around two hundreds of polyploids have been reported in insect and vertebrate [24,25,26,27,28]. To reduce errors and ensure the authenticity and reliability of data, it is necessary to choose the most appropriate reference genes for different ploidy cells or organisms
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