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Role and Gonadotrophic Regulation of X-Linked: MATERIALS AND METHODS(1)

METHODS(1)

Materials

Culture media and reagents were purchased from Gibco Bethesda Research Laboratories (Burlington, ON, Canada). Agarose (low gelling temperature), DNA (calf thymus), Hoechst 33258 compound, Triton X-100, Tween 20, collagenase (type 1A), DNase 1, RNase A, bovine insulin, human transferrin, ascorbic acid, sodium selenite anhydrous, and eCG were obtained from Sigma Chemical Company (St. Louis, MO). The enhanced chemiluminescence (ECL) detection kit, [methyl-3H]thymidine (25 Ci/mmol), and [a-32P]ddATP were obtained from Amersham Life Science (Oakville, ON, Canada). Ovine FSH (NIAMDD oFSH-14) was obtained from the National Institute of Diabetes & Digestive & Kidney Diseases (Baltimore, MD). canadian pharmacy family viagra

Acrylamide (electrophoresis grade), A^-methylene-bis-acrylamide, ammonium persulfate, glycine, SDS-PAGE prestained molecular weight standards (low range), nitrocellulose membranes, and horseradish peroxidase (HRP)-conjugated anti-rabbit and anti-mouse immunoglobulin (Ig) G were products from Bio-Rad (Richmond, CA). Rhoda-mine-conjugated goat anti-rabbit IgG was a product of Santa Cruz Biotechnology, Inc. (Santa Cruz, CA). Mounting medium for fluorescence was purchased from Vector Laboratories, Inc. (Vectashield, H-1000; Burlingame, CA). Cell death detection (TUNEL) kit and Terminal End Transferase (TdT) were purchased from Boehringer-Mannhein (Montreal, PQ, Canada) and Alamar Blue dye from Biosource Incorporated (distributor: Medicorp, Montreal, PQ, Canada). The Qiagen Tissue Amp Kit and QIA-quick Nucleotide Removal Kit were products of Qiagen (Qiagen, Inc., Chatsworth, CA). The IM-8 Micro-injector and Joystick (mN-151) as well as Micromanipulator were purchased from Narishige International, Inc. (Long Island, NY). The stage warmer was bought from Linkam Scientific Instruments (Surry, U.K.).