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Reference gene selection for expression studies in the reproductive axis tissues of Magang geese at different reproductive stages under light treatment
In quantitative PCR research, appropriate reference genes are key to determining accurate mRNA expression levels. In order to screen the reference genes suitable for detecting gene expression in tissues of the reproductive axis, a total of 420 (males and females = 1:5) 3-year-old Magang geese were selected and subjected to light treatment. The hypothalamus, pituitary and testicular tissues were subsequently collected at different stages. Ten genes including
HPRT1
,
GAPDH
,
ACTB
,
LDHA
,
SDHA
,
B2M
,
TUBB4
,
TFRC
,
RPS2
and
RPL4
were selected as candidate reference genes. The expression of these genes in goose reproductive axis tissues was detected by real-time fluorescent quantitative PCR. The ΔCT, geNorm, NormFinder and BestKeeper algorithms were applied to sort gene expression according to stability. The results showed that
ACTB
and
TUBB4
were the most suitable reference genes for the hypothalamic tissue of Magang goose in the three breeding stages;
HPRT1
and
RPL4
for pituitary tissue; and
HPRT1
and
LDHA
for testicular tissue. For all three reproductive axis tissues,
ACTB
was the most suitable reference gene, whereas the least stable reference gene was
GAPDH
. Altogether, these results can provide references for tissue expression studies in geese under light treatment.