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Sep 25, 2014;7 675.

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

Jae-Kyo Jeong, Min-Hee Kang, Sangiliyandi Gurunathan, Ssang-Goo Cho, Chankyu Park, Han Geuk Seo, Jin-Hoi Kim
Author Information
  1. Jin-Hoi Kim: Department of Animal Biotechnology, KonKuk University, Seoul 143-701, Republic of Korea. jhkim541@konkuk.ac.kr.
PMID: 25256308 DOI: 10.1186/1756-0500-7-675

Abstract

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.

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MeSH Term

Animals
Blastocyst
Culture Media
Databases, Genetic
Embryo Culture Techniques
Female
Gene Expression Profiling
Gene Expression Regulation, Developmental
Genetic Markers
Male
Mice
Mice, Inbred C57BL
Mice, Inbred ICR
RNA Stability
RNA, Messenger
Real-Time Polymerase Chain Reaction
Reference Standards
Reproducibility of Results
Reverse Transcriptase Polymerase Chain Reaction
Species Specificity

Chemicals

Culture Media
Genetic Markers
RNA, Messenger
Comparative Study Journal Article

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