Evaluation of reference genes in mouse preimplantation embryos for gene expression studies using real-time quantitative RT-PCR (RT-qPCR)

BMC Research Notes

Jin-Hoi Kim¹  Han Geuk Seo¹  Chankyu Park¹  Ssang-Goo Cho¹  Sangiliyandi Gurunathan¹  Min-Hee Kang¹  Jae-Kyo Jeong¹ 

[1] Department of Animal Biotechnology, KonKuk University, Seoul 143-701, Republic of Korea

关键词: Gene expression;    Preimplantation embryos;    Mouse;    Reference gene;    RT-PCR;   

Others  :  1127420
DOI  :  10.1186/1756-0500-7-675

 received in 2014-03-09, accepted in 2014-09-05,  发布年份 2014

【 摘 要 】

Background

Real-time quantitative reverse-transcriptase polymerase chain reaction (RT-qPCR) is the most sensitive, and valuable technique for rare mRNA detection. However, the expression profiles of reference genes under different experimental conditions, such as different mouse strains, developmental stage, and culture conditions have been poorly studied.

Results

mRNA stability of the actb, gapdh, sdha, ablim, ywhaz, sptbn, h2afz, tgfb1, 18 s and wrnip genes was analyzed. Using the NormFinder program, the most stable genes are as follows: h2afz for the B6D2F-1 and C57BL/6 strains; sptbn for ICR; h2afz for KOSOM and CZB cultures of B6D2F-1 and C57BL/6 strain-derived embryos; wrnip for M16 culture of B6D2F-1 and C57BL/6 strain-derived embryos; ywhaz, tgfb1, 18 s, 18 s, ywhaz, and h2afz for zygote, 2-cell, 4-cell, 8-cell, molular, and blastocyst embryonic stages cultured in KSOM medium, respectively; h2afz, wrnip, wrnip, h2afz, ywhaz, and ablim for zygote, 2-cell, 4-cell, 8-cell, molular, and blastocyst stage embryos cultured in CZB medium, respectively; 18 s, h2afz, h2afz, actb, h2afz, and wrnip for zygote, 2-cell, 4-cell, 8-cell, molular, and blastocyst stage embryos cultured in M16 medium, respectively.

Conclusions

These results demonstrated that candidate reference genes for normalization of target gene expression using RT-qPCR should be selected according to mouse strains, developmental stage, and culture conditions.

【 授权许可】

   
2014 Jeong et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

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