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Reference gene selection for gene expression study in tissues of long-tailed chickens KCI 등재
- 언어KOR
- URLhttps://db.koreascholar.com/Article/Detail/404386
pp.152-164
Quantitative real-time polymerase chain reaction (RTqPCR) is a rapid and precise method of analysis to quantify the level of gene expression and is widely used in the diagnosis of diseases and quantitative analysis of genes. In RT-qPCR analysis, a reference gene (or housekeeping gene) is used for normalization of experimental results. Since this method of analysis detects a small quantity of the product, it is highly sensitive and it is important for the accuracy and reproducibility of the experiment to select a reference gene suitable for gene expression studies. As the expression levels of the reference gene are affected under different conditions, in order to determine the suitability of the housekeeping gene used as the reference gene, it is necessary to verify the expression stability. In the current study, the stability of the expression of 11 housekeeping genes (B2M, SDHA, GAPDH, RPL13, VIM, EEF1A1, HPRT1, GUSB, RPL19, ACTB, and ABL1) was investigated in the tissues of long-tailed chickens (heart, thigh, and breast). Expression stability evaluation was analyzed with four software: BestKeeper, NormFinder, geNorm, and RefFinder. In our study, GAPDH in heart tissue, HPRT1 in thigh tissue, and RPL13 in breast tissue were selected as the most stable reference genes. Evaluation of the expression stability of housekeeping genes can provide important data in gene expression studies by selecting an appropriate reference gene according to various conditions.
Materials and Methods
RNA 추출법
하우스키핑 유전자의 프라이머 제작
cDNA 합성
Real-time quantitative PCR (RT-qPCR) 분석
하우스키핑 유전자의 발현 안정성 분석
Results
PCR에서 하우스키핑 유전자들의 발현
Bestkeeper 분석
NormFinder 분석
RefFidner 분석
Discussion
References
