Name: rnai-mediated gene knockdown and in vivo diuresis assay in adult female aedes aegypti mosquitoes
Uploaded: 2012-07-14T14:00:00
Description: Watch this Scientific Journal Video about RNAi-mediated Gene Knockdown and In Vivo Diuresis Assay in Adult Female Aedes aegypti Mosquitoes at JoVE.com

Die Autoren danken Victoria Carpenter für ihre kritischen Kommentare dieses Protokolls.

Teil I - RNAi-vermittelten Gen-Knockdown in der Erwachsenenbildung Aedes aegypti Mücken. Für Experiment Überblick siehe Abbildung 1. 1. dsRNA-Synthese <ol><li> Synthetisieren spezifische dsRNAs gegen Gene von Interesse und Kontrolle dsRNAs. Anmerkung: Es wird empfohlen die Entwicklung Primer für die PCR-Fragmente im Bereich von 300 bis 500 Basenpaare am 3&#39;-Ende der spezifischen cDNA 2 und mit dem T7-Primer-Sequenz am 5 &#39;befestigt Ende (5&#39;-T...

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Die RNAi-Protokoll verwendet wurde im Labor von Alexander Raikhel wurde an der University of California Riverside 6,7 entwickelt und ist vergleichbar mit einem Protokoll, das von Garver und Dimopoulos 4 veröffentlicht. Der experimentelle Ansatz in diesem Video-Protokoll gezeigt werden verwendet, um Gene in Insektenzellen Diurese in einem in-vivo-Umgebung verwickelt zu werden. Die Ausscheidungsorgane der Insekten, die Malpighischen Gefäßen, haben das Interesse von Generationen v...

<table border="1"><tbody><tr><td> Name des Reagenz oder Ausrüstung </td><td> Firma </td><td> Katalog-Nummer </td><td> Kommentare </td></tr><tr><td> MEGAscript T7 High-Yield-Kit </td><td> Ambion, Inc. </td><td> AM1334 </td><td></td></tr><tr><td> PBS-Puffer </td><td> Sigma-Aldrich </td><td> P4417 </td><td></td></tr><tr><td> Kunststoffschlauch </td><td> Lokale Anbieter </td><td></td><td> PVC </td></tr><tr><td> 1 ml Kunststoff-Pipettenspitze </td><td> VWR </td><td> 83007-376 </td><td> Blaue Spitze </td></tr><tr><td> 1 ml-Spritze </td><td> Becton, Dickinson and Company </td><td> 309602 </td><td></td></tr><tr><td> Schere </td><td> Lokale Anbieter </td><td></td><td></td></tr><tr><td> Metall-Nadel </td><td> Carolina Biologicals </td><td> 654307 </td&gt; <td> Größe 5 </td></tr><tr><td> Fly-Pad </td><td> Genesee Scientific </td><td> 789060 </td><td></td></tr><tr><td> Batteriebetriebene Aspirator w / Sammelfläschchen </td><td> Upma Labs </td><td> IPMM 2000 </td><td></td></tr><tr><td> Feine Spitze Pinzette </td><td> World Precision Instruments </td><td> 14095 </td><td></td></tr><tr><td> Glaskapillare Nadeln </td><td> World Precision Instruments </td><td> 1B200-6 </td><td></td></tr><tr><td> Stereo Dissektionsmikroskop </td><td> Leica Microsystems </td><td> S6D </td><td></td></tr><tr><td> Analytische Präzisionswaage </td><td> Mettler Toledo </td><td> AB54S </td><td></td></tr><tr><td> Saccharose </td><td> Sigma-Aldrich </td><td> 84097 </td><td></td></tr><tr><td> Ein Pint gewachst ausgekleideten Karton Tassen </td><td> Lokale Anbieter </td><td></td><td> Hergestellt Suppentassen </td></tr><tr><td> Mesh-Netz </td><td> Lokale Anbieter </td><td></td><td> Kunststoff-fly Gaze </td></tr></tbody></table>

rnai-mediated gene knockdown and in vivo diuresis assay in adult female aedes aegypti mosquitoes

Dieses Video demonstriert Protokoll eine effektive Technik, um ein bestimmtes Gen in einer Insekten und die Durchführung einer neuartigen Bioassay zur Ausscheidung zu messen. Diese Methode kann verwendet werden, um ein besseres Verständnis des Prozesses der Diurese bei Insekten erhalten werden und ist besonders nützlich bei der Untersuchung der Diurese in blutsaugende Arthropoden, die aufnehmen können große Mengen an Flüssigkeit in einem einzigen Blutmahlzeit sind. Das RNAi-vermittelten Gen Knockdown mit einem In-vivo-Assay kombiniert Diurese wurde von der Hansen-Labor entwickelt, um die Effekte der RNAi-vermittelte Reduktion von Aquaporin-Gene auf Aedes aegypti Moskito Diurese 1 studierst. Das Protokoll ist Setup in zwei Teile: der erste Beispiel demonstriert, wie man ein einfaches Moskitonetz Injektionsgerät konstruieren und zur Vorbereitung und injizieren dsRNA in den Brustkorb von Mücken für RNAi-vermittelte Gen-Knockdown. Das zweite Beispiel demonstriert, wie Sie feststellen,Ausscheidung in Moskitos mit einem In-vivo-Bioassay.

Watch this Scientific Journal Video about RNAi-mediated Gene Knockdown and In Vivo Diuresis Assay in Adult Female Aedes aegypti Mosquitoes at JoVE.com

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

In diesem Protokoll kombinieren wir RNAi-vermittelte Gen-Silencing mit ein<em&gt; In-vivo-</em&gt; Diurese Assay, um die Auswirkungen Knockdown von Genen von Interesse Studie wurde auf Moskito-Flüssigkeit ausgeschieden.

RNAi-vermittelte Gen-Knockdown und<em&gt; In Vivo</em&gt; Diurese Assay in Adult Female<em&gt; Aedes aegypti</em&gt; Moskitos

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

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

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