Screening and Stability Analysis of Reference Genes for Gene Expression Normalization in Hybrid Yellow Catfish (Pelteobagrus fulvidraco ♀ × Pelteobagrus vachelli ♂) Fed Diets Containing Different Soybean Meal Levels

In this study, we screened the expression stability of six reference genes (18S rRNA, β-actin, GAPDH, EF1a, B2M, and HPRT1) in hybrid yellow catfish (n = 6), considering SBM levels, sampling time points, different tissues. Four statistical programs, BestKeeper, NormFinder, Genorm, Delta Ct, combined with a method that comprehensively considered all results, were used to evaluate these systematically. The results showed levels significantly impacted most studied impact was time-, dose-, tissue-dependent. varied depending on tissue, point, dosage. Additionally, more variations found among tissues than or points. Due its high expression, 18S rRNA excluded from list candidate genes. β-actin GAPDH liver HPRT1 EF1a intestine stable when considered. HPRT1, sampled at 2 W 4 6 proposed as two When points considered, top three intestine. contrast, B2M are liver. summary, study. depends tissues, diet catfish. Therefore, attention should be paid factors before selecting suitable for normalizing target