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Reference gene selection for real-time quantitative PCR analysis on ovarian cryopreservation by vitrification in mice
Shan Yuanyuan1, Su Qin, Xu Rongrong
1Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education, Key Laboratory of Reproduction and Genetics in Ningxia, Department of Biochemistry and Molecular Biology, Ningxia Medical University, Yinchuan, Ningxia, 75004, People's Republic of China.
Purpose:
To ensure the correct interpretation of the results of quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) from ovarian tissue cryopreserved by vitrification, it is critical to normalize expression levels to a reference gene with stable messenger RNA (mRNA) expression in the vitrified/warmed ovarian tissue. The aim of this work was to identify suitable reference genes for qRT-PCR analysis during ovarian cryopreservation by vitrification.
Methods:
GeNorm, NormFinder, comparative Delta-CT, and BestKeeper were used to analyze the expression and stability of the 14 reference genes GAPDH, ABL1, ACTB, CDKN1A, GPER, GUSB, HPRT1, HSP90AB1, IPO8, PPIA, RPL4, RPL30, TBP, and UPAR.
Results:
Our results indicated that ACTB and RPL4 were relatively stable reference genes in vitrified/warmed ovaries.
