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

Forfatterne takker Victoria Carpenter for hennes kritiske kommentarer av denne protokollen.

Del I - RNAi-mediert genet knockdown i voksen Aedes aegypti mygg. For eksperiment oversikt se Figur 1. 1. dsRNA Synthesis <ol><li> Syntetisere spesifikke dsRNAs mot gener av interesse og kontroll dsRNAs. Merk: Vi anbefaler utviklingsland primere for PCR fragmenter i området 300 til 500 basepar er plassert på 3 &#39;enden av spesifikke cDNA 2 og med T7 primer sekvensen festet på 5&#39; enden (5&#39;-TAA TAC GAC TCA CTA TAG GG-3 &#39;). Unikhet av fragmen...

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Den RNAi protokollen som brukes er utviklet i laboratoriet av Alexander Raikhel ved University of California Riverside 6,7 og ligner på en protokoll publisert av Garver og Dimopoulos 4. Den eksperimentelle tilnærmingen vist i denne videoen protokollen kan brukes til å studere gener involvert i insekt diurese i en in vivo setting. De ekskresjonsfunksjoner organer insekter, de Malpighian tubuli, har tiltrukket seg interesse av generasjoner av forskere som en &quot;enkel&quot; modell syste...

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

Denne videoen protokollen demonstrerer en effektiv teknikk for å knockdown et bestemt gen i en insekt og gjennomføre en roman bioassay å måle utskillelse rate. Denne metoden kan brukes til å oppnå en bedre forståelse av prosessen diurese i insekter og er spesielt nyttig i studiet av diuresen i blod som ammer leddyr som er i stand til å ta opp store mengder væske i et enkelt måltid blod. Dette RNAi-mediert genet knockdown kombinert med en in vivo diurese analysen ble utviklet av Hansen lab for å studere effektene av RNAi-mediert knockdown av aquaporin gener på Aedes aegypti mygg diurese en. Protokollen er oppsettet i to deler: den første demonstrasjonen illustrerer hvordan å konstruere en enkel mygg injeksjon enhet og hvordan man forbereder og injisere dsRNA inn thorax av mygg for RNAi-mediert genet knockdown. Den andre demonstrasjonen viser hvordan du finnerutskillelse priser i mygg ved hjelp av en 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

I denne protokollen kombinerer vi RNAi-mediert genet Silencing med en<em&gt; In vivo</em&gt; Diurese analysen for å studere effekter knockdown av gener av interesse har på mygg væske utskillelse.

RNAi-mediert Gene knockdown og<em&gt; In Vivo</em&gt; Diurese Assay i voksen hunn<em&gt; Aedes aegypti</em&gt; Mygg

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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This report describes a step-by-step guide to the technique of acute intrathecal needle injections in a noninvasive manner, i.e. independent of catheter ...

Fat Body Organ Culture System in Aedes Aegypti, a Vector of Zika Virus

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Aedes aegypti mosquitoes experimentally infected with Wolbachia are being utilized in programs to control the spread of arboviruses such as dengue, ...

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Forbedret fecundity og ufrivillig analyse for Aedes aegypti ved hjelp av 24 Brønn vev kulturplater (EAgaL plater)

Mosquitoes represent a significant public health problem as vectors of various pathogens. For those studies that require an assessment of mosquito fitness ...