title: Evaluation of reference genes in mouse preimplantation embryos for gene expression studies using real-time quantitative RT-PCR (RT-qPCR).
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.
authors.forename: Jae-Kyo
authors.initials: JK
authors.lastname: Jeong
authors.forename: Min-Hee
authors.initials: MH
authors.lastname: Kang
authors.forename: Sangiliyandi
authors.initials: S
authors.lastname: Gurunathan
authors.forename: Ssang-Goo
authors.initials: SG
authors.lastname: Cho
authors.forename: Chankyu
authors.initials: C
authors.lastname: Park
authors.forename: Han Geuk
authors.initials: HG
authors.lastname: Seo
authors.affiliation: Department of Animal Biotechnology, KonKuk University, Seoul 143-701, Republic of Korea. jhkim541@konkuk.ac.kr.
authors.forename: Jin-Hoi
authors.initials: JH
authors.lastname: Kim
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000378:metabolism*; D003470:Culture Media; D030541:Databases, Genetic; D046149:Embryo Culture Techniques; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods* / Q000592:standards; D018507:Gene Expression Regulation, Developmental; D005819:Genetic Markers; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D020871:RNA Stability; D012333:RNA, Messenger / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards; D012015:Reference Standards; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000592:standards; D013045:Species Specificity
chemical_list: D003470:Culture Media; D005819:Genetic Markers; D012333:RNA, Messenger