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

Gli autori ringraziano Victoria Carpenter per i suoi commenti critici di questo protocollo.

Parte I - RNAi-mediato knockdown gene in adulti zanzare Aedes aegypti. Esperimento per una panoramica si veda la Figura 1. 1. dsRNA Sintesi <ol><li> Sintetizzare dsRNA specifici contro i geni di dsRNA di interesse e di controllo. Nota: Si consiglia di primer di sviluppo per frammenti PCR nell&#39;intervallo di 300 a 500 coppie di basi situate all&#39;estremità 3 &#39;del cDNA specifico 2 e con la sequenza del primer T7 attaccata all&#39;estremità 5&#39; ...

<ol>
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Il protocollo utilizzato RNAi è stato sviluppato nel laboratorio di Alexander Raikhel presso la University of California Riverside 6,7 ed è simile a un protocollo pubblicato da Garver e Dimopoulos 4. L&#39;approccio sperimentale mostrato in questo protocollo video possono essere utilizzati per studiare i geni coinvolti nella diuresi insetto in un ambiente in vivo. Gli organi escretori di insetti, i tubuli malpighiani, hanno attirato l&#39;interesse di generazioni di ricercatori come un s...

<table border="1"><tbody><tr><td> Nome del reagente o attrezzature </td><td> Azienda </td><td> Numero di catalogo </td><td> Comments </td></tr><tr><td> MEGAscript T7 Kit High Yield </td><td> Ambion, Inc. </td><td> AM1334 </td><td></td></tr><tr><td> Tampone PBS </td><td> Sigma-Aldrich </td><td> P4417 </td><td></td></tr><tr><td> Tubi in plastica </td><td> Rivenditore locale </td><td></td><td> PVC </td></tr><tr><td> 1 ml di plastica della punta di pipetta </td><td> VWR </td><td> 83007-376 </td><td> Blu punta </td></tr><tr><td> 1 ml siringa </td><td> Becton, Dickinson and Company </td><td> 309602 </td><td></td></tr><tr><td> Forbici </td><td> Rivenditore locale </td><td></td><td></td></tr><tr><td> Metallo ago </td><td> Carolina Biologicals </td><td> 654307 </td&gt; <td> Size 5 </td></tr><tr><td> Vola pad </td><td> Genesee Scientific </td><td> 789060 </td><td></td></tr><tr><td> Alimentato a batteria Aspiratore w / collezione vial </td><td> Upma Labs </td><td> IPMM 2000 </td><td></td></tr><tr><td> Pinza punta fine </td><td> Mondo Strumenti di precisione </td><td> 14095 </td><td></td></tr><tr><td> Aghi di vetro capillari </td><td> Mondo Strumenti di precisione </td><td> 1B200-6 </td><td></td></tr><tr><td> Dissezione microscopio stereo </td><td> Leica Microsystems </td><td> S6D </td><td></td></tr><tr><td> Bilancia analitica di precisione </td><td> Mettler Toledo </td><td> AB54S </td><td></td></tr><tr><td> Saccarosio </td><td> Sigma-Aldrich </td><td> 84097 </td><td></td></tr><tr><td> Una pinta di cartone cerato tazze allineate </td><td> Rivenditore locale </td><td></td><td> Tazze di zuppa di Prodotto </td></tr><tr><td> Delle maglie delle reti </td><td> Rivenditore locale </td><td></td><td> fly garza di plastica </td></tr></tbody></table>

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

Questo protocollo video mostra una tecnica efficace per abbattere un particolare gene in un insetto e condurre un biotest romanzo per misurare il tasso di escrezione. Questo metodo può essere utilizzato per ottenere una migliore comprensione del processo di diuresi in insetti ed è particolarmente utile nello studio della diuresi nel sangue alimentazione artropodi in grado di raccogliere grandi quantità di liquido in un pasto di sangue singolo. Questo RNAi-mediato knockdown gene combinato con un test in vivo diuresi è stato sviluppato dal laboratorio di Hansen per studiare gli effetti di RNAi-mediato knockdown di geni acquaporina sulla diuresi zanzara Aedes aegypti 1. Il protocollo è configurato in due parti: la prima dimostrazione illustra come costruire un semplice dispositivo di iniezione zanzare e su come preparare ed iniettare dsRNA nel torace di zanzare per RNAi-mediato knockdown gene. La seconda dimostrazione illustra come determinaretassi di escrezione nelle zanzare utilizzando un saggio biologico in vivo.

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 questo protocollo si combinano RNAi-mediato silenziamento genico con un<em&gt; In vivo</em&gt; Test diuresi per studiare il colpo degli effetti dei geni di interesse ha sulla escrezione di liquidi zanzara.

RNAi-mediato Knockdown Gene e<em&gt; In Vivo</em&gt; Saggio La diuresi in femmina adulta<em&gt; Aedes aegypti</em&gt; Le zanzare

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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