https://wiki.xenbase.org/xenwiki/index.php?title=In_vitro_fertilization_of_Xenopus_laevis_(Conlon_lab)&feed=atom&action=historyIn vitro fertilization of Xenopus laevis (Conlon lab) - Revision history2024-03-29T14:29:19ZRevision history for this page on the wikiMediaWiki 1.40.1https://wiki.xenbase.org/xenwiki/index.php?title=In_vitro_fertilization_of_Xenopus_laevis_(Conlon_lab)&diff=47320&oldid=previmported>Virgilio: Created page with "Protocol submitted by VGP from Conlon lab protocols [http://www.unc.edu/~fconlon/Protocols.htm#tropprot] link to protocol document [http://www.unc.edu/~fconlon/Protocols/in%20v..."2011-11-22T14:50:31Z<p>Created page with "Protocol submitted by VGP from Conlon lab protocols [http://www.unc.edu/~fconlon/Protocols.htm#tropprot] link to protocol document [http://www.unc.edu/~fconlon/Protocols/in%20v..."</p>
<p><b>New page</b></p><div>Protocol submitted by VGP from Conlon lab protocols [http://www.unc.edu/~fconlon/Protocols.htm#tropprot]<br />
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link to protocol document [http://www.unc.edu/~fconlon/Protocols/in%20vitro%20fertilization.doc]<br />
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in vitro Fertilization of Xenopus laevis Protocol<br />
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Modified 4/15/2005<br />
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Egg Collection<br />
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1) Allow female to lay eggs in 1X MMR<br />
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2) Collect eggs in a small beaker in 1X MMR<br />
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Testis Preparation<br />
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1) Remove testis from male into 1X MBS<br />
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2) Remove blood, fat, and debris while in 1X MBS<br />
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3) Place testis in L15 media<br />
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4) Wash testis in 1X MBS before mincing (Perform all of the following as quickly as possible)<br />
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5) Mince testis with razorblade on a petri dish cover<br />
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Fertilization<br />
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1) Remove all but 20 ml of 1X MMR from beaker with eggs<br />
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2) Quickly add 40 ml 0.1X MBS to beaker (making it roughly 33% MMR)<br />
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3) Let eggs settle and quickly remove as much liquid as possible<br />
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4) Add 1ml 0.1X MBS to testis and resuspend sperm<br />
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5) Add the 1ml of sperm to the beaker with the eggs<br />
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6) Swirl a few times (I actually swirl the eggs and sperm once every few minutes until fertilized)<br />
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7) The eggs should fertilize and rotate within 10-15 minutes @ RT or 20-30 min @ 17°C</div>imported>Virgilio