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Dejellying embryos (Zorn lab) - Revision history
2024-03-29T07:30:26Z
Revision history for this page on the wiki
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https://wiki.xenbase.org/xenwiki/index.php?title=Dejellying_embryos_(Zorn_lab)&diff=47329&oldid=prev
205.142.197.33 at 15:01, 7 December 2011
2011-12-07T15:01:23Z
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<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Older revision</td>
<td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Revision as of 09:01, 7 December 2011</td>
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<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>| </div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>|<ins style="font-weight: bold; text-decoration: none;">to pH 8.0 </ins></div></td></tr>
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<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>|</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>|<ins style="font-weight: bold; text-decoration: none;">to pH 8.0</ins></div></td></tr>
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<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|200 mls </div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>|200 mls </div></td></tr>
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<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>|</div></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>|<ins style="font-weight: bold; text-decoration: none;">to pH 8.0</ins></div></td></tr>
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<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div><del style="font-weight: bold; text-decoration: none;">Volume of 0.1xMBS Cysteine 5N NaOH</del></div></td><td colspan="2" class="diff-side-added"></td></tr>
<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div><del style="font-weight: bold; text-decoration: none;">50 mls 1g</del></div></td><td colspan="2" class="diff-side-added"></td></tr>
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<tr><td class="diff-marker" data-marker="−"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div><del style="font-weight: bold; text-decoration: none;">200 mls 4g</del></div></td><td colspan="2" class="diff-side-added"></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><br/></td></tr>
<tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Dissolve the Cysteine hydrochloride (or L-Cysteine hydrochloride-1-hydrate) in ~3/4 the final volume of 0.1xMBS. </div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Dissolve the Cysteine hydrochloride (or L-Cysteine hydrochloride-1-hydrate) in ~3/4 the final volume of 0.1xMBS. </div></td></tr>
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205.142.197.33
https://wiki.xenbase.org/xenwiki/index.php?title=Dejellying_embryos_(Zorn_lab)&diff=47328&oldid=prev
imported>Virgilio: Created page with "Protocol submitted by VGP Dejellying solution: 2% Cysteine-hydrochloride, pH 8 in 0.1xMBS, to remove the jelly coat from fertilised Xenopus eggs and embryos before they can be..."
2011-11-22T19:45:36Z
<p>Created page with "Protocol submitted by VGP Dejellying solution: 2% Cysteine-hydrochloride, pH 8 in 0.1xMBS, to remove the jelly coat from fertilised Xenopus eggs and embryos before they can be..."</p>
<p><b>New page</b></p><div>Protocol submitted by VGP<br />
<br />
<br />
Dejellying solution:<br />
<br />
2% Cysteine-hydrochloride, pH 8 in 0.1xMBS, to remove the jelly coat from fertilised Xenopus eggs and embryos before they can be manipulated. <br />
<br />
{|border="1" cellpadding="5"<br />
|-<br />
! scope="col" | Volume of 0.1xMBS<br />
! scope="col" | Cysteine<br />
! scope="col" | 5N NaOH<br />
|-<br />
|50 mls<br />
|1g<br />
| <br />
|-<br />
|100 mls<br />
|2g<br />
|<br />
|-<br />
|200 mls <br />
|4g<br />
|<br />
|}<br />
<br />
Volume of 0.1xMBS Cysteine 5N NaOH<br />
50 mls 1g<br />
100 mls 2g<br />
200 mls 4g<br />
<br />
Dissolve the Cysteine hydrochloride (or L-Cysteine hydrochloride-1-hydrate) in ~3/4 the final volume of 0.1xMBS. <br />
<br />
Adjust to ~ pH 8 with 5N NaOH. <br />
<br />
Check that the pH is correct and then adjust the final volume with 0.1xMBS. <br />
<br />
This solution will only last one day.<br />
<br />
<br />
Dejelling embryos:<br />
<br />
Wait at least one hour after fertilisation, any time after this embryos may be dejellied. <br />
<br />
Decant the MBS off the embryos, leaving them semi-dry and stuck to the bottom of the dish. <br />
<br />
Cover the embryos in the 2% cysteine solution, which will dissolve the jelly coats surrounding the eggs. <br />
<br />
With occasional, mild agitation, the embryos will be liberated in 5-7 minutes. <br />
<br />
It is important not to “swirl” eggs too vigorously when dejellying as this can cause axis duplications in some cases. <br />
<br />
Once the jelly coat is dissolved, the cysteine solution must be removed immediately (it can be reused). <br />
<br />
The embryos are then washed 3-4 times in a large volume of 0.1xMBS to remove residual cysteine. <br />
<br />
If left in the cysteine solution too long, or not washed enough, the plasma membrane will be damaged and the embryos will die!</div>
imported>Virgilio