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Biological Characteristics and Data (Mice)

Mouse Surgery Protocol ( Click to Download ) Mouse Surgery Protocols (edited 1/11/96) A. Mice, like most species have a circadian rhythm. Investigators should be aware that this may affect biological data and should standardize the time of day that samples/measurements are taken to avoid this affect. The standard light/dark cycle at the University of […]

DMBA Treatment

Mouse Surgery Protocol ( Click to Download ) Mouse Surgery Protocols (edited 1/11/96) A. Dimethylbenzanthracene (DMBA, Calbiochem) will be dissolved in cottonseed oil to yield a 0.25% (w/v/) solution. A dose of 0.2 ml will be administered by gastric intubation once each week for 3 consecutive weeks starting when the mice are 8 weeks old. […]

Embryo Transfer and Screening Transgenic Mice

Mouse Surgery Protocol ( Click to Download ) Mouse Surgery Protocols (edited 1/11/96) A. Oocytes for pronuclear injection are collected from females that have been hormonally stimulated to cause superovulation. The eggs are flushed from the oviducts following euthanasia of the donor. After pronuclear injection of the DNA, the embryos are surgically injected into the […]

Hormone BrdU Aministsration

hormone_BrDU ( Click to Download ) •Bed the animals on SaniChips, rather than corn pellets, to prevent any interference from phytoestrogens for 5-7 days before the hormone treatment. •Two days before sacrifice, inject mice with 100 μl of E+P under the skin between the shoulder blades. [The final dose given is 1 μg E2 and […]

Hormone Treatment of Mice

Mouse Surgery Protocol ( Click to Download ) Mouse Surgery Protocols (edited 1/11/96) A. Estrus cycle and hormone profiles Vaginal smears (method and stages from Boot, Kwa and Ropcke, 1981 in Mammary Tumors in the Mouse, Hilgers & Sluyser, Eds.; see ref #2679) – A cotton swab is used to collect cells from the vagina […]

Irradiation of Mice

Mouse Surgery Protocol ( Click to Download ) Mouse Surgery Protocols (edited 1/11/96) A. Gamma-irradiation will be administered at the dose rate of 400 Rads/minute using 137Cs source located in Room 112 of Paige laboratory. Total whole body doses will not exceed 500 Rad. This will be accomplished by placing single mice in a cardboard […]

Mouse Estrous Cycle

Stage Ovary Uterus Vagina Smear Diestrus Small follicles only are present with large corpora lutea from the previous ovulation. These secrete for only a very short time unless pregnancy or pseudopregnancy intervene. Small and anaemic, low motility, lumen small and slit-like. Cells of the uterine mucosa columnar; polymorphonuclear leucocytes in stroma; endometrial glands collapsed, atrophic. […]

Protocol For Genotyping ERKO Mice

Oligos: UY83 (P1); UV182 (P2); WR115 (P3); WR111 (P4) P1 5′ TTG CCG ATA ACA ATA ACA T 3′ P2 5′ ATT GTC TCT TTC TGA CAC 3′ P3 5′ GGC ATT ACC ACT TCT CCT GGG AGT CT 3′ P4 5′ TCG CTT TCC TGA AGA CCT TTC ATA T 3′ PCR Mixes (for […]

Cell line Xenograft Injections (PJ Keller)

Xenograft Injection Prep ( Click to Download ) Preparing cells for injection: Determine the total volume of injections (plan on 40 µl per gland plus enough for 1-2 extra glands per cell line/condition). Thaw out enough matrigel on ice for 50% of the total volume of injections. Trypsinize cells to a single cells suspension and […]