Get A Laboratory Guide to the Mammalian Embryo PDF

By David K. Gardner, Michelle Lane, Andrew J. Watson

ISBN-10: 0195142268

ISBN-13: 9780195142266

This ebook pulls jointly the total diversity of mobilephone tradition, biochemical, microscopic, and genetic concepts to review the early mammalian embryo. formerly, there hasn't ever been this type of entire compendium, even though there were extra concentrated books of protocol, similar to Manipulating the Mouse Embryo, from chilly Spring Harbor. This publication is meant to entice all constituencies, from easy experimental technological know-how to scientific and animal technological know-how functions.

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Extra resources for A Laboratory Guide to the Mammalian Embryo

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3. 5°C. Leave the tubes for 5 min to allow the cumulus-enclosed oocyte complexes to settle in the bottom of the tube. 4. Locate cumulus-enclosed oocytes from the follicular aspirate. A grid is scratched onto the outer bottom of a 10-mm petri dish using a scalpel blade or needle to assist in the searching. Using a sterile Pasteur pipette, remove the bottom 1 ml of the follicular aspirate containing the sediment from the tube and transfer it to the searching plate. To this sediment add 1–2 ml of HEPES-TCM-199 medium.

Discard the oviduct and transfer a new oviduct to the collection dish. 1). 5. Add the 500 ml of warmed hyaluronidase solution (1 mg/ml, 1100 U/ml) to the dish to remove cumulus cells. Expose for 20–60 s and collect embryos as soon as they become denuded. 6. During the exposure of the cumulus complex to hyaluronidase, set up three washing drops of handling medium (approximately 50 ml) in a collection dish or the lid of a collection dish. 7. Wash denuded embryos through the three drops of handling medium.

Spray the back of the mouse with 70% ethanol. 3. Using forceps, make a part in the hair of the mouse ventrally down the midline of the back. This part should be around 3–4 cm long and adjacent to the top of the hips of the mouse. 4. Make an incision along the part through the skin to expose the body cavity. The middle of this incision should be adjacent to the hips of the mouse. This will ensure that the reproductive tract is easy to find. Wipe the incision with 70% ethanol to remove any loose hairs.

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A Laboratory Guide to the Mammalian Embryo by David K. Gardner, Michelle Lane, Andrew J. Watson

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