Download PDF by David K. Gardner, Michelle Lane, Andrew J. Watson: A Laboratory Guide to the Mammalian Embryo
By David K. Gardner, Michelle Lane, Andrew J. Watson
This publication pulls jointly the complete diversity of mobilephone tradition, biochemical, microscopic, and genetic suggestions to check the early mammalian embryo. formerly, there hasn't ever been this kind of finished compendium, even though there were extra centred books of protocol, corresponding to Manipulating the Mouse Embryo, from chilly Spring Harbor. This ebook is meant to entice all constituencies, from easy experimental technological know-how to scientific and animal technology purposes.
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Extra resources for A Laboratory Guide to the Mammalian Embryo
Embryo transfer Preparation of pseudopregnant recipients Ideally females should be 8–12 weeks of age. 1. Place two mature females with the vasectomized males the evening before the desired day 1 of pregnancy. The number of plugs can be increased by assessing the stage of the estrous cycle by the appearance of the vagina (20). 2. The following morning assess mating by the presence of a vaginal plug. This is designated day 1 of pseudopregnancy. Transfer media Embryos should be transferred in a HEPES/MOPS modification of the culture medium, as the time taken to load and transfer the embryos would result in a significant loss in CO2 from bicarbonate-buffered medium and a rise in pH.
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7. Wash denuded embryos through the three drops of handling medium. 8. Embryos are now ready to be placed into culture. Embryos from some strains are very susceptible to their environment. Therefore, it is important to minimize the time from extraction of the cumulus masses and placement in culture. It is advisable to keep this time period to a maximum of 10 min. Therefore, it is preferable to do multiple smaller embryo collections rather than one large collection. 1. Collection of mouse zygotes from the ampullary region of the oviduct: (a) The swollen ampulla containing the cumulus-oocyte complex is located.
A Laboratory Guide to the Mammalian Embryo by David K. Gardner, Michelle Lane, Andrew J. Watson