Final oocyte maturation is the key step to successful spawning and fertilization.Quantitative real-time PCR（q PCR） is the technique of election to quantify the abundance of functional genes in such study. Reference ...Final oocyte maturation is the key step to successful spawning and fertilization.Quantitative real-time PCR（q PCR） is the technique of election to quantify the abundance of functional genes in such study. Reference gene is essential for correct interpretation of q PCR data. However, an ideal universal reference gene that is stable under all experimental circumstances has not been described. Researchers should validate their reference genes while performing q PCR analysis. The expression of 6 candidate reference genes： 18 s r RNA,28 s r RNA, Cathepsin Z, Elongation factor 1-α, Glyceraldehyde-3-phosphate dehydrogenase andβ-actin were investigated during final oocyte maturation induced by different compounds（DES and DEHP） in common carp（Cyprinus carpio）. Four softwares（Bestkeeper, ge Norm,Norm Finder and Ref Finder） were used to screen the most stable gene in order to evaluate their expression stability. The results revealed that EF1α was highly stable expressed when final oocyte maturation was induced by DES, while gapdh was the most stable gene when final oocyte maturation was induced by DEHP. Stable expressed reference gene selection is critical for all q PCR analysis to get accurate target gene m RNA expression information.展开更多
基金supported by the Innovation Team Fundation of Jimei University(No.2010A001)
文摘Final oocyte maturation is the key step to successful spawning and fertilization.Quantitative real-time PCR（q PCR） is the technique of election to quantify the abundance of functional genes in such study. Reference gene is essential for correct interpretation of q PCR data. However, an ideal universal reference gene that is stable under all experimental circumstances has not been described. Researchers should validate their reference genes while performing q PCR analysis. The expression of 6 candidate reference genes： 18 s r RNA,28 s r RNA, Cathepsin Z, Elongation factor 1-α, Glyceraldehyde-3-phosphate dehydrogenase andβ-actin were investigated during final oocyte maturation induced by different compounds（DES and DEHP） in common carp（Cyprinus carpio）. Four softwares（Bestkeeper, ge Norm,Norm Finder and Ref Finder） were used to screen the most stable gene in order to evaluate their expression stability. The results revealed that EF1α was highly stable expressed when final oocyte maturation was induced by DES, while gapdh was the most stable gene when final oocyte maturation was induced by DEHP. Stable expressed reference gene selection is critical for all q PCR analysis to get accurate target gene m RNA expression information.
