Screening of Stably Expressed Internal Reference Genes for Quantitative Real-Time PCR Analysis in Quail

Yuan, Zhiwen; Zhang, X. H.; Pang, You Zhi; Qi, Yanxia; Wang, Q. K.; Ren, S. W.; Hu, Yuxin; Zhao, Yongtao; Wang, T.; Huo, Liang-xiao

doi:10.1134/s1062359022050223

Cited by 6 publications

(3 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Four statistical algorithms, GeNorm, NormFinder, BestKeeper and ∆Ct, were used to evaluate the stability of these selected reference genes [32]. The most stable reference genes screened from several statistical algorithms were different, while the least stable reference gene was highly consistent.…”

Section: Discussionmentioning

confidence: 99%

Selection of Reliable Reference Genes for Gene Expression Normalization in Sagittaria trifolia

Tang

Liu

et al. 2023

Genes

View full text Add to dashboard Cite

Real-time quantitative PCR (RT-qPCR) is a method with high sensitivity and convenience that has been extensively used to analyze the expression level of target genes. A reference gene with a highly stable expression is required to ensure the accuracy of experimental results. However, the report on appropriate reference genes in arrowheads (Sagittaria trifolia) is still limited. In this study, eight candidate reference genes (ACT5, UBQ, GAPDH, CYP, NAC, IDH, SLEEPER and PLA) were selected. The candidate genes were employed in a RT-qPCR assay in different tissues at different developmental stages of the same tissue (including corm, leaf and leafstalk) in arrowheads. Five statistical algorithms, GeNorm, NormFinder, BestKeeper, delta cycle threshold (ΔCt) and RefFinder, were used to evaluate the stability of these genes’ expressions in order to identify the appropriate reference genes. The results showed that UBQ was the optimum reference gene in leaf, leafstalk, root, stolon and corm, IDH exhibited the most stable expression during the expansion of corm, UBQ and PLA were the most stable reference genes in developmental stages of leaf and leafstalk, respectively. Finally, the reliability of reference genes was further confirmed by the normalization of PDS and EXP1 genes under different arrowhead tissues and developmental stages of corm, respectively. This study constitutes important guidance for the selection of reliable reference genes for analyzing the tissue- and developmental-stage-specific expression of genes in arrowheads.

show abstract

Section: Discussionmentioning

confidence: 99%

Selection of Reliable Reference Genes for Gene Expression Normalization in Sagittaria trifolia

Tang

Liu

et al. 2023

Genes

View full text Add to dashboard Cite

show abstract

“…Using eukaryotic translation initiation factor 2 subunit 3 ( EIF2S3 )[ 12 ] as the internal reference gene, RT-qPCR was used to detect the expression levels of the SLC45A2 gene in E10 embryos of Beijing white and Chinese yellow quail. SLC45A2 and EIF2S3 primers were designed using Primer 5.0 software ( Table 1 ),which were synthesised by Beijing Tsingke Biotech Co., Ltd.…”

Section: Methodsmentioning

confidence: 99%

Expression and Mutation of <i>SLC45A2</i> Affects Iris Color in Quail

Huo,

Zhang,

Pang

et al. 2024

J. Poult. Sci.

View full text Add to dashboard Cite

“…Primers were designed using primer 5.0 software (Table 1). EIF2S3 was used as internal control, and the

2^{- ΔΔ c_{t}}

method was used to calculate the fold change of gene relative expression (Yuan et al., 2022).…”

Section: Methodsmentioning

confidence: 99%

RNA sequencing analysis reveals that missense mutation in SOX10 is associated with iris color phenotype in quail

Ren,

Zhang,

Pang

et al. 2023

Animal Genetics

View full text Add to dashboard Cite

To investigate the molecular mechanisms underlying the differences in iris color in quail, the transcriptome of iris tissue from black quail and Korean quail at day 10 of hatching was RNA sequenced in this study. The differentially expressed genes (DEGs) were screened, functionally annotated and enriched after the quality control and mapping of the raw data. RT‐qPCR validation was performed using EIF2S3 as an internal reference gene. The screened SNPs were studied by bioinformatics analysis and iris color correlation analysis. The results showed that there were 425 upregulated genes and 364 downregulated genes in 789 DEGs. Gene Ontology (GO) enrichment analysis revealed that 139 DEGs were significantly enriched in 154 GO terms. The Kyoto Encyclopedia of Genes and Genomes enrichment results showed that the Notch signaling pathway, melanogenesis and tyrosine metabolism were associated with pigment synthesis (p < 0.05). The expression levels of the ASIP, MLPH, PMEL, TYR and SOX10 genes were significantly different in black quail iris and Korean quail iris, as verified by RT‐qPCR. The SOX10 gene c.324G>C mutation, which caused the replacement of p.Glu108Asp, had a highly significant correlation with iris color in black quail and Korean quail, which may be one of the reasons for different in iris color between these two quail species.

show abstract

Screening of Stably Expressed Internal Reference Genes for Quantitative Real-Time PCR Analysis in Quail

Cited by 6 publications

References 34 publications

Selection of Reliable Reference Genes for Gene Expression Normalization in Sagittaria trifolia

Selection of Reliable Reference Genes for Gene Expression Normalization in Sagittaria trifolia

Expression and Mutation of <i>SLC45A2</i> Affects Iris Color in Quail

RNA sequencing analysis reveals that missense mutation in SOX10 is associated with iris color phenotype in quail

Contact Info

Product

Resources

About