Name: microinjection of a. aegypti embryos to obtain transgenic mosquitoes
Uploaded: 2007-07-04T00:00:00
Description: Watch this Scientific Journal Video about Microinjection of A. aegypti Embryos to Obtain Transgenic Mosquitoes at JoVE.com

<p class="jove_title"> Voorbereiding van de micro-injectie </p><ol><li> Bloed te voeden muggen: voor injectie van maandag tot vrijdag voeden vrouwen op de vorige donderdag. </li><li> Bereid glas naalden met behulp van het programma 3 of 2 (zie ook Materiaal). </li><li> Leggen buis voor de embryo&#39;s is Drosophila cultuur flacon. Natte watten in de bodem, met een natte schijf van filtreerpapier te bedekken. </li><li> Bereid plastic dekglas door steken dubbelzijdig tape om het ene uiteinde. Trim tape om dekglas, zodat het eindigt aan de rand va...

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		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>1x Injection solution</td>
<td>Buffer</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>5mM KCl, 0.1mM sodium phosphate, ph 6. 8</td>
</tr>
<tr>
<td>Construct plasmid</td>
<td>DNA</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>0.5 mg/ml.  </td>
</tr>
<tr>
<td>Helper plasmid</td>
<td>DNA</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>0.3 mg/ml </td>
</tr>
<tr>
<td>EndoFree Plasmid kit</td>
<td>Kit</td>
<td>Qiagen</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Millex-GV column</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>SLGV RO4 NL</td>
<td>To clean DNA.</td>
</tr>
<tr>
<td>Glass puller</td>
<td>Instrument</td>
<td>Sutter Instrument Co</td>
<td>Model P-87</td>
<td>Heat-815, Pull-10, Vel-10, Time-100.
Heat-700, Pull-30, Vel-36, Time-50
</td>
</tr>
<tr>
<td>Quartz puller</td>
<td>Instrument</td>
<td>Sutter Instrument Co</td>
<td>Model P-2000</td>
<td>Heat-275, Fil-3, Vel-38, Del-250, Pull-141</td>
</tr>
<tr>
<td>Injection glass needles</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Prepared by drawing capillary tubing into a fine 3-8 um with a shaft of approximately 100-300 um micropipette puller. When the needle is new, the tip is sealed and usually breaks in the first or second injection or the glass needle tip can be beveled using a Brown type micro-pipette beveller (Sutter Instrument).</td>
</tr>
<tr>
<td>Aedes aegypti</td>
<td>Animal</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Male and female mosquitos, to obtain embryos.</td>
</tr>
<tr>
<td>Drosophila culture vial</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>For mosquito egg-laying.</td>
</tr>
<tr>
<td>Halocarbon oil</td>
<td>Reagent</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Oil for desiccation: Halocarbon oil 700:Halocarbon oil 27(1:1)</td>
</tr>
<tr>
<td>Forceps No5</td>
<td>Tool</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Fine paintbrush</td>
<td>&nbsp;</td>
<td>Sable</td>
<td>No. 0000</td>
<td>&nbsp;</td>
<tr>
<td>Whatman paper</td>
<td>Tool</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Microloader</td>
<td>Tool</td>
<td>Eppendorf</td>
<td>5242 956.003</td>
<td>&nbsp;</td>
<tr>
<td>Transjector</td>
<td>Tool</td>
<td>Eppendorf</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Microscope</td>
<td>&nbsp;</td>
<td>Leica</td>
<td>&nbsp;</td>
<td>With moving stage and micromanipulator.</td>
</tr>
<tr>
<td>3MM Whatman paper</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>double sided tape</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>
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microinjection of a. aegypti embryos to obtain transgenic mosquitoes

<p class="jove_content">In deze video, Nijole Jasinskiene toont de methoden die gebruikt worden om transgene Aedes aegypti muggen, die vectoren voor knokkelkoorts te genereren. De technieken voor het correct opstellen van micro-injectie naalden, droogmiddelen embryo&#39;s en het uitvoeren van micro-injectie worden aangetoond.</p>

Watch this Scientific Journal Video about Microinjection of A. aegypti Embryos to Obtain Transgenic Mosquitoes at JoVE.com

Microinjection of A. aegypti Embryos to Obtain Transgenic Mosquitoes

Micro-injectie van A. aegypti embryo&#39;s om transgene muggen verkrijgen

In this video, Nijole Jasinskiene demonstrates the methodology employed to generate transgenic Aedes aegypti mosquitoes, which are vectors for dengue ...

Research

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Protocol for Dengue Infections in Mosquitoes (A. aegypti) and Infection Phenotype Determination

Protocol for Dengue Infections in Mosquitoes A. aegypti and Infection Phenotype Determination

Protocol voor Dengue Infecties in Muggen (A. aegypti) en Infectie Fenotype Bepaling

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Dissection of Midgut and Salivary Glands from Ae. aegypti Mosquitoes

Ontleding van Midgut en speekselklieren van de Ae. aegypti Muggen

The mosquito midgut and salivary glands are key entry and exit points for pathogens such as Plasmodium parasites and Dengue viruses. This video protocol ...

Building a Better Mosquito: Identifying the Genes Enabling Malaria and Dengue Fever Resistance in A. gambiae and A. aegypti Mosquitoes

Bouwen aan een betere Mosquito: Het identificeren van de genen Enabling malaria en knokkelkoorts verzet in A. gambiae en A. aegypti muggen

In this interview, George Dimopoulos focuses on the physiological mechanisms used by mosquitoes to combat Plasmodium falciparum and dengue virus ...

Generation of Stable Transgenic C. elegans Using Microinjection

Generatie van Stable transgene C. elegans Met behulp van micro-injectie

Transgenic Caenorhabditis elegans can be readily created via microinjection of a DNA plasmid solution into the gonad 1.  The plasmid DNA rearranges to ...

Microinjection of mRNA and Morpholino Antisense Oligonucleotides in Zebrafish Embryos.

Micro-injectie van mRNA en morfolino antisense oligonucleotiden in zebravis embryo&#39;s.

An essential tool for investigating the role of a gene during development is the ability to perform gene knockdown, overexpression, and misexpression ...

Microinjection of Zebrafish Embryos to Analyze Gene Function

Micro-injectie van de zebravis embryo&#39;s Gene functie analyseren

One of the advantages of studying zebrafish is the ease and speed of manipulating protein levels in the embryo.  Morpholinos, which are synthetic ...

Microinjection of Medaka Embryos for use as a Model Genetic Organism

Micro-injectie van Medaka embryo&#39;s voor gebruik als een model organisme Genetische

In this video, we demonstrate the technique of microinjection into one-cell stage medaka embryos. Medaka is a small egg-laying freshwater fish that allows ...

RNAi-mediated Gene Knockdown and <em>In Vivo</em> Diuresis Assay in Adult Female <em>Aedes aegypti</em> Mosquitoes

RNAi-mediated Gene Knockdown and In Vivo Diuresis Assay in Adult Female Aedes aegypti Mosquitoes

RNAi-gemedieerde Gene Knockdown en<em&gt; In Vivo</em&gt; Urineproductie Assay in het volwassenenonderwijs Vrouw<em&gt; Aedes aegypti</em&gt; Muggen

This video protocol demonstrates an effective technique to knockdown a particular gene in an insect and conduct a novel bioassay to measure excretion ...

Generation of Transgenic <em>Hydra</em> by Embryo Microinjection

Generation of Transgenic Hydra by Embryo Microinjection

Generatie van transgene<em&gt; Hydra</em&gt; Door Embryo Microinjection

As a member of the phylum Cnidaria, the sister group to all bilaterians, Hydra can shed light on fundamental biological processes shared among ...