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

Yazarlar bu protokolün onun eleştirel yorumlar için Victoria Carpenter teşekkür ederim.

Bölüm I - yetişkin Aedes aegypti sivrisinekler RNAi-aracılı gen devirme. Deney genel bakış için Şekil 1&#39;e bakınız. 1. dsRNA Sentezi <ol><li> Ilgi ve kontrol dsRNAs genler karşı spesifik dsRNAs sentezlerler. Not: 3 &#39;özgü cDNA 2 sonu ve 5 de ekli T7 primer sekans ile&#39; de uç (5&#39;-GAC TAA TAC TCA CTA bulunan 300-500 baz çifti aralığında PCR parçaları için gelişmekte olan primerler önermektedir TAG GG-3 &#39;). Parçalar?...

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Kullanılan RNAi protokolü California Riverside 6,7 Üniversitesi&#39;nde Alexander Raikhel laboratuvarında geliştirilen ve Garver ve Dimopoulos 4 tarafından yayınlanan bir protokol benzer olmuştur. Bu video protokolü gösterildiği deneysel yaklaşım in vivo ortamda bir böcek diürez ilgili genler incelemek için kullanılabilir. Böcekler, Malpighi tüpleri tübüller, ve boşaltım organları diürez için &#39;basit&#39; bir model sistem olarak araştırmacıların nesillerin ...

<table border="1"><tbody><tr><td> Reaktif ya da ekipman Adı </td><td> Şirket </td><td> Katalog numarası </td><td> Yorumlar </td></tr><tr><td> MEGAscript T7 Yüksek Kapasiteli Kiti </td><td> Ambion, Inc </td><td> AM1334 </td><td></td></tr><tr><td> PBS tamponu </td><td> Sigma-Aldrich </td><td> P4417 </td><td></td></tr><tr><td> Plastik boru </td><td> Yerel satıcı </td><td></td><td> PVC </td></tr><tr><td> 1 ml plastik pipet </td><td> VWR </td><td> 83007-376 </td><td> Mavi ucu </td></tr><tr><td> 1 ml şırınga </td><td> Becton, Dickinson and Company </td><td> 309602 </td><td></td></tr><tr><td> Makas </td><td> Yerel satıcı </td><td></td><td></td></tr><tr><td> Metal iğne </td><td> Carolina Biologicals </td><td> 654307 </td&gt; <td> Boyut 5 </td></tr><tr><td> Pad Fly </td><td> Genesee Bilimsel </td><td> 789060 </td><td></td></tr><tr><td> Pille aspiratör w / tahsilat flakon </td><td> UPMA Labs </td><td> ESYM 2000 </td><td></td></tr><tr><td> İnce uçlu forseps </td><td> Dünya Hassas Aletler </td><td> 14095 </td><td></td></tr><tr><td> Cam kılcal iğneler </td><td> Dünya Hassas Aletler </td><td> 1B200-6 </td><td></td></tr><tr><td> Stereo diseksiyon mikroskobu </td><td> Leica Microsystems </td><td> S6D </td><td></td></tr><tr><td> Analitik hassas denge </td><td> Mettler Toledo </td><td> AB54S </td><td></td></tr><tr><td> Sakaroz </td><td> Sigma-Aldrich </td><td> 84097 </td><td></td></tr><tr><td> Bir bardak bira kaplı karton bardak mumlu </td><td> Yerel satıcı </td><td></td><td> Üretici çorba bardak </td></tr><tr><td> Mesh net </td><td> Yerel satıcı </td><td></td><td> plastik sinek gazlı bez </td></tr></tbody></table>

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

Bu video protokolü bir böcek belirli bir gen devirme ve atılım hızı ölçmek için yeni bir biyoassay yapmak için etkili bir teknik gösterilmiştir. Bu yöntem, böceklerin diürez işlemi daha iyi anlaşılmasını sağlamak için kullanılan ve tek bir kan unu sıvı büyük miktarlarda kadar alabilmesi için olan bir kan-besleme eklem bacaklıların in diürez çalışmada özellikle yararlıdır edilebilir. In vivo diürez tayininde birlikte bu RNAi-aracılı gen Knockdown Aedes aegypti sivrisinek diürez 1 aquaporin genlerin RNAi aracılı Knockdown etkilerini incelemek için Hansen laboratuar tarafından geliştirilmiştir. Protokol iki bölüm halinde kurulum: ilk gösteri basit bir sivrisinek enjeksiyon cihazı nasıl oluşturacağını ve RNAi-aracılı gen devirme için sivrisinek toraks içine dsRNA hazırlamak ve enjekte nasıl göstermektedir. İkinci gösteri nasıl belirleneceği gösterilmektedirin vivo biyoassay bir kullanarak sivrisinekler atılımı oranları.

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

Bu protokolü biz bir ile RNAi-aracılı gen susturulması birleştirmek<em&gt; In vivo</emIlgi genlerin etkilerini demonte çalışma&gt; diürez testi sivrisinek sıvı atılımı vardır.

RNAi-aracılı Gene Nakavt ve<em&gt; In Vivo</emYetişkin Kadınlar&gt; diürez Testi<em&gt; Aedes aegypti</em&gt; Sivrisinekler

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

Research

JoVE Journal

Biology

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

Injection of dsRNA into Female A. aegypti Mosquitos

Bayan A. aegypti Sivrisinek içine dsRNA enjeksiyon

Reverse genetic approaches have proven extremely useful for determining  which genes underly resistance to vector pathogens in mosquitoes.  This video ...

Microinjection of A. aegypti Embryos to Obtain Transgenic Mosquitoes

Transgenik Sivrisinekler Edinme A. aegypti Embriyolar Mikroenjeksiyon

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

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

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

Dang Enfeksiyonlar Sivrisinekler (A. aegypti) ve Enfeksiyon Fenotip belirlenmesi için protokol

The purpose of this procedure is to infect the Aedes mosquito with dengue virus in a laboratory condition and examine the infection level and dynamic of ...

Dissection of Midgut and Salivary Glands from Ae. aegypti Mosquitoes

Ae midgut ve Tükürük Bezlerinin Diseksiyon. aegypti Sivrisinekler

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

RNAi Mediated Gene Knockdown and Transgenesis by Microinjection in the Necromenic Nematode Pristionchus pacificus

RNAi Mediated Gene Knockdown and Transgenesis by Microinjection in the Necromenic Nematode Pristionchus pacificus

RNAi Necromenic nematodu Mikroenjeksiyon Gen demonte ve Transgenesis aracılığı ile Pristionchus pacificus</em

Although it is increasingly affordable for emerging model organisms to obtain completely sequenced genomes, further in-depth gene function and expression ...

Using SecM Arrest Sequence as a Tool to Isolate Ribosome Bound Polypeptides

Ribozom Ciltli Polipeptitler izole bir aracı olarak SECM Tutuklanma sırası kullanılarak

Extensive research has provided ample evidences suggesting that protein folding in the cell is a co-translational process1-5. However, the exact pathway ...

In Vivo SiRNA Transfection and Gene Knockdown in Spinal Cord via Rapid Noninvasive Lumbar Intrathecal Injections in Mice

In Vivo SiRNA Transfection and Gene Knockdown in Spinal Cord via Rapid Noninvasive Lumbar Intrathecal Injections in Mice

Farelerde Hızlı noninvaziv Lomber İntratekal enjeksiyon yoluyla Vivo siRNA&#39;nın Transfaksiyon ve Omurilik Gen demonte olarak

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

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

Yağ vücut Organ kültür sistemi içinde Aedes Aegypti, bir vektör Zika virüs

The insect fat body plays a central role in insect metabolism and nutrient storage, mirroring functions of the liver and fat tissue in vertebrates. Insect ...

Maintaining Aedes aegypti Mosquitoes Infected with Wolbachia

Maintaining Aedes aegypti Mosquitoes Infected with Wolbachia

Aedes aegypti sivrisinek enfekte Wolbachia ile bakımını yapma

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

Improved Fecundity and Fertility Assay for Aedes aegypti using 24 Well Tissue Culture Plates (EAgaL Plates)

Improved Fecundity and Fertility Assay for Aedes aegypti using 24 Well Tissue Culture Plates EAgaL Plates

24 Kuyu Doku Kültürü Plakası (EAgaL Plakaları) kullanılarak Aedes aegypti için Geliştirilmiş Doğurganlık ve Doğurganlık Tahlilleri

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