Upregulation of the BRCA1 gene in human germ cells and in preimplantation embryos  Sandrine Giscard d’Estaing, Ph.D., Delphine Perrin, M.Sc., Gilbert M. Lenoir, Ph.D., Jean François Guérin, Ph.D., Robert Dante, Ph.D.  Fertility and Sterility  Volume 84, Issue 3, Pages 785-788 (September 2005) DOI: 10.1016/j.fertnstert.2005.02.037 Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

FIGURE 1 Reverse transcriptase polymerase chain reaction assay of BRCA1 mRNA. (A) Representative RT-PCR assay of BRCA1 RNAs. Ten microliters of RT-PCR reaction mixture (total volume 100 μL), performed from a fixed amount of RNA mixed with known amounts of competitor BRCA1 RNA, were analyzed on a 2% agarose gel. Filled squares, 258 base pair (bp) wild-type BRCA1 band; open squares, 306-bp competitor BRCA1 band. Diagrammatic representation of the above data; the normalized signals corresponding to the target mRNA and the competitor were plotted against the initial number of competitor molecules added to the test tubes. The abscissa of the intersection of the curves represents an estimation of the equivalence point between the initial amount of the competitor molecules and the number of copies of the mRNA assayed. The RT-PCR reactions were performed with six oocytes per point: Lane 1, 5 × 103; Lane 2, 10 × 103; Lane 3, 50 × 103 of competitor molecules. (B) Number of BRCA1 transcripts. BRCA1 mRNA copies per microgram of RNA: blue bars, immature male germ cells, 11 × 106; pink bars, oocytes, 4 × 106; purple bars, blastocysts, 0.7 × 106; green bars, granulosa cells, 0.17 × 106. Giscard d’Estaing. BRCA1 expression in human germ cells. Fertil Steril 2005. Fertility and Sterility 2005 84, 785-788DOI: (10.1016/j.fertnstert.2005.02.037) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions