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

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

This publication pulls jointly the complete diversity of phone tradition, biochemical, microscopic, and genetic suggestions to check the early mammalian embryo. previously, there hasn't ever been this type of finished compendium, notwithstanding there were extra targeted books of protocol, similar to Manipulating the Mouse Embryo, from chilly Spring Harbor. This ebook is meant to attract all constituencies, from easy experimental technology to scientific and animal technological know-how applications.

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Repeat by inserting the needle into the other uterine horn and flush medium through this horn. 6. Collect and wash embryos in another well of handling medium. 5. 3) to 37°C. Attach a sterile 27-gauge needle to 1-ml syringe. Prepare pulled Pasteur pipettes with an internal volume of about 500 mm for manipulation of the cumulus-enclosed oocytes complexes. A pipette that is just larger than the complexes is essential to ensure adequate washing and minimal transfer of medium between drops. Procedure 1.

6. When all of the antral follicles have been aspirated, collect the cumulus-oocyte complexes using the pulled Pasteur pipette and place them into a fresh dish of HEPES maturation medium. 6. Maturation of mouse germinal vesicle stage oocytes Preparation of Maturation dishes Germinal vesicle stage oocytes are matured overnight for approximately 16–17 h at 37°C in 6% CO2:5%O2:89%N2. PREPARATION OF GAMETES, IVM, IVF, AND EMBRYO RECOVERY 33 The morning of culture, dishes containing maturation medium are prepared.

10. Using watchmaker’s forceps, grab the reproductive tract at the utero-tubal junction. Pick up a 27-G needle attached to a 1-ml syringe and also the pipette with the embryos. 11. Pull the uterus taut. 12. Using the 27-G needle, make a hole in the uterus around one-third of the way down from the oviduct. Ensure that the hole goes into the lumen of the uterus and not into the muscle walls. Remove the needle and put aside while keeping your eye on the hole. 13. Insert the pipette into the hole made with the needle.

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