His offspring have won 143 graded races, including 28 Grade I races and 4 Nippon Derbys in Japan.
The ZnO thin films were deposited on a glass (OA-10G, Nippon Electronic Glass Co., Ltd., Otsu, Shiga, Japan) substrate using the described RF magnetron sputtering system.
Cells were first acclimated in RPMI 1640 or DMEM containing 5% FBS for 16 h before exposure to GUT-70 (Nippon Shinyaku, Kyoto, Japan) (Kimura et al, 2005).
Both the unprocessed and digested PCR products were subjected to electrophoresis in 3% agarose gel (Agarose 21, Nippon Gene Co., Ltd., Tokyo, Japan).
One microliter of the SLiCE reaction solution was chemically transformed into 20 μL ECOS X Competent E. coli DH5α (Nippon gene, Tokyo, Japan) according to the instruction manual.
ECOS X Competent E. coli DH5α (Nippon gene, Tokyo, Japan) chemically competent cells were used to transform the recombinant DNA generated using the SLiCE method, in order to obtain the constant transformation efficiency.
Confocal images were acquired with a Zeiss Observer Z.1 inverted microscope (Carl Zeiss Microimaging GmbH, Goettingen, Germany) equipped with a Yokogawa CSU-X1 Nipkow spinning disk system (via Visitron Systems GmbH, Puchheim, Germany), a piezoelectric z-axis motorized stage (CRWG3-200, Nippon Thompson Co., Ltd, Tokyo, Japan), and a CoolSNAP HQ2 CCD Camera (Photometrics, Tucson, AZ).
MAS1 siRNA (siRNA) or control scramble siRNA (10 pmol, encapsulated with cationic liposome [EL-C-01; Nippon-Oil & Fats, Tokyo, Japan]), 21, 22 CDDP (3 mg/kg body weight, once a week) and XTN (0.5 mg/kg body weight, once a week) 23 were injected into the peritoneal cavity.
The TP53 gene, exon 5 to exon 9 including exon intron junctions, were amplified by polymerase chain reaction (PCR) using "p53 primers" (Nippon Gene) and Ex Taq DNA polymerase with 3′ exonuclease activity (TaKaRa Bio Inc., Tokyo, Japan).
High rates of cleavage blocking and low rates of embryo production are both common in in vitro fertilization studies on ruminants, particularly among wild species (Gazella dama mhorr[ 62], Cervus nippon and Cervus elaphus[ 65]), and these factors seem to be related to inadequate medias for development in vitro[ 65- 68].
