Name: in ovo electroporations of hh stage 10 chicken embryos
Uploaded: 2007-11-01T00:29:01
Description: Watch this Scientific Journal Video about In Ovo Electroporations of HH Stage 10 Chicken Embryos at JoVE.com

<p class="jove_step">蛋处理</p><ol><li>鸡蛋可能保持在13 ° C，无生存能力的重大损失（是用于此目的的理想选择的一个小酒柜）一个星期前，以孵化。 </li><li>孵化之前，让鸡蛋温暖至室温，然后放置在加湿的鸡孵化器设置到37.8℃（100℉）。 </li><li>我们通常electroporate胚胎大约48小时，从孵化开始，当胚胎已达到汉堡包和汉密尔顿阶段10。 </li><li>根据所需的汉堡包和汉密尔顿阶段，培养时间可能会有所不同。温度是至关重...

<ol>
	<li>Itasaki, N., Bel-Vialar, S., Krumlauf, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=&apos;Shocking&apos;+developments+in+chick+embryology:+electroporation+and+in+ovo+gene+expression.">&apos;Shocking&apos; developments in chick embryology: electroporation and in ovo gene expression.</a> <em style='font-style: italic'>Nat Cell Biol</em>. <b>1</b>, 203-207 (1999).</li></ol>

<p>该协议提供了一个分步指南HH10鸡胚胎神经管电。除了显示技术，还提供了存储和处理的鸡蛋准则。这种技术具有广泛的应用范围为遗传分析和实验的易用性和低成本使它们成为许多实验室是可行的。 </p>

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
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<tr>
<td>Chicken egg incubator</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>humidified, set to 37.80&deg;C (100F).  </td>
</tr>
<tr>
<td>Pulse generator </td>
<td>&nbsp;</td>
<td>Genetronics </td>
<td>BTX T820 </td>
<td>Square wave generator or comparable</td>
</tr>
<tr>
<td>Electrodes</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>our electrodes were made from 0.25 mm diameter platinum wire and were 5 mm long and spaced 1 mm apart</td>
</tr>
<tr>
<td>Connecter cables</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Micromanipulator</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Dissecting scope </td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>with large working distance</td>
</tr>
<tr>
<td>Glass capillaries </td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>and capillary puller or commercial glass needles</td>
</tr>
<tr>
<td>Leibovitz&#x2019;s L-15 media</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>chicken eggs</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>fertilized</td>
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<tr>
<td>Curved scissors</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>10 ml syringe</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Large bore needle </td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>16-18G</td>
</tr>
<tr>
<td>Mouth pipette</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>India ink </td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Insulin syringe/syringe </td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Plasmid DNA </td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&#x2265; 1&#181;g/&#181;l in sterile TE or 1X PBS containing 0.05% Fast Green dye</td>
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in ovo electroporations of hh stage 10 chicken embryos

<p class="jove_content">庞大的规模和外部发展的鸡胚早已在发育生物学研究的一个有价值的工具。随着分子生物学技术的问世，小鸡已经成为一个有用的系统来研究基因表达调控和功能。通过DNA或RNA的结构到发展中国家的鸡胚电穿孔，基因表达或撞倒在体内的基因功能分析。同样，记者构造可用于映射的命运，或检查公认的基因调控元件。小鸡电相类似的实验，在小鼠，具有快捷，方便和价格低廉的优势。第一视频演示如何使一个窗口，在蛋壳操纵胚胎。下一步，胚胎是用下面的胚胎注入印度墨水的稀溶液中观察。一个玻璃针和吸管是用来注入发展中国家的神经管，铂电极平行放置胚胎，短的电脉冲脉冲发生器的管理DNA和快速的绿色染料。最后，鸡蛋是用胶带密封，并放入孵化器的进一步发展。此外，视频显示正确的鸡蛋储存和处理，并讨论可能的原因，胚胎丢失以下电。</p>

Watch this Scientific Journal Video about In Ovo Electroporations of HH Stage 10 Chicken Embryos at JoVE.com

In Ovo Electroporations of HH Stage 10 Chicken Embryos

<p class="jove_content">OVO电小鸡是一种技术，这使得禽流胚胎的基因操纵。这项技术的常见应用包括基因的功能分析和公认的增强子元件。本视频演示了为HH 10鸡胚神经管电。注射技术和适当的鸡蛋处理进行了讨论。</p>

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