受害者还是个中国人(GUO),这次这个女的真的完了
http://stapcells.blogspot.jp/2014/02/blog-post_7834.html"Stimulus-triggered fate conversion of somatic cells into pluripotency"
Nature 505, 641–647 (30 January 2014)
著者: Haruko Obokata (小保方晴子), Teruhiko Wakayama (若山照彦), Yoshiki Sasai (笹井芳樹), Koji Kojima (小島宏司), Martin P. Vacanti (マーティン・バカンティ), Hitoshi Niwa (丹羽仁史), Masayuki Yamato (大和雅之), Charles A. Vacanti (チャールズ・バカンティ)
Karyotype analysis
Karyotype analysis was performed by Multicolor FISH analysis (M-FISH). Subconfluent STAP stem cells were arrested in metaphase by colcemid (final concentration 0.270 μg ml−1) to the culture medium for 2.5 h at 37 °C in 5% CO2. Cells were washed with PBS, treated with trypsin and EDTA (EDTA), re-suspended into cell medium and centrifuged for 5 min at 1,200 r.p.m. To the cell pellet in 3 ml of PBS, 7 ml of a pre-warmed hypotonic 0.0375 M KC1 solution was added. Cells were incubated for 20 min at 37 °C. Cells were centrifuged for 5 min at 1,200 r.p.m. and the pellet was re-suspended in 3–5 ml of 0.0375 M KC1 solution. The cells were fixed with methanol/acetic acid (3:1; vol/vol) by gently pipetting. Fixation was performed four times before spreading the cells on glass slides.
[source]
Multicolor karyotype analyses of mouse embryonic stem cells.
In Vitro Cell Dev Biol Anim. 2005 Sep-Oct;41(8-9):278-83.
Guo J1, Jauch A, Heidi HG, Schoell B, Erz D, Schrank M, Janssen JW.
1Institute of Human Genetics, University of Heidelberg, D-69120 Heidelberg, Germany.
Materials and Methods
Chromosome preparation
Metaphase spreads of the ES cells were performed as follows. Subconfluent ES cells were arrested in metaphase by adding colcemid (final concentration 0.270 μg/ml) to the culture medium for 2.5 h at 37° C in 5% CO2. Cells were washed with PBS, treated with trypsin-ethylenediaminetetraacetic acid (EDTA), resuspended into cell medium and centrifuged for 5 min at 1200 rpm. To the cell pellet in 3 ml of PBS, 7 ml of a prewarmed hypotonic 0.0375 M KCl solution was added. Cells were incubated for 20 min at 37° C. Cells were centrifuged for 5 min at 1200 rpm and the pellet was resuspended in 3–5 ml of 0.0375 M KCl solution. The cells were fixed with methanol/acetic acid (3:1, vol:vol) by gently pipetting. Fixation was performed four times prior to spreading the cells on glass slides.