البعوض الغذائية : <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> الغذاء ج : أسس الغذاء الأسماك (Aquaricare). ألف قرص صغير يحتاج إلى إضافته. </li><li style=";text-align:right;direction:rtl"> الغذائية ب : أسس الغذاء لجنة مناهضة التعذيب (بورينا). ألف قرص صغير يحتاج إلى إضافته. </li><li style=";text-align:right;direction:rtl"> الغذاء C : القط الغذاء (بورينا). قرصين يحتاج إلى إضافته. </li></ol> الدم : ويمكن استخدام الحيوانات أو البعوض إلى دم الإنسان الخلفي. ويتم جمع الدم الطازج مع حقنة وضعت في أنبوب معقم 15 مل تحتوي على فالكون 1ml من CPD (تخثر) لكل 10 مل من الدم ، مختلطة ثم بلطف وطرد في درجة حرارة الغرفة على 2000 دورة في الدقيقة لمدة 5 دقائق. يتم تجاهل طاف ، مع الحرص على إزالة الطبقة الشهباء الذي يضم خلايا الدم الأخرى (مثل WBC). ومن ثم علق بيليه RBC في حجم مساو من قبل pipetting RPMI المتوسطة ، وغسلها أخرى في هذا ثلاث مرات المتوسط. بعد غسل النهائي ، ومعلق في كرات الدم الحمراء في مكعبات حجم مساو من RPMI المتوسطة وتخزينها في 4 درجات مئوية (يمكن أن تبقى لمدة 8-10 أيام). قبل الرضاعة ، وطرد من ربك (في RPMI) (2000 دورة في الدقيقة لمدة 5 دقائق) لتكوير أسفل الخلايا ، ومعلق في RBC معبأة في المصل (O + مصل بشري) للحصول على الهيماتوكريت 40 ٪. يجب أن يبقى في الدم عند 37 درجة مئوية دائما للتغذية. DAY 1 : <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> تتغذى على البعوض 3-5 اليوم البالغين من العمر الإناث على الدم لوضع البيض. </li><li style=";text-align:right;direction:rtl"> لتغذية الدم ، يتم استخدام طريقة مصطنعة غشاء التغذية (parafilm) على النحو التالي : <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> تختلط خلايا الدم الحمراء (انظر أعلاه لإعداد الدم) مع المصل الحرارة المعطل للحصول على الهيماتوكريت ~ 40 ٪ (40 ٪ معبأة RBC ومصل الدم 60 ٪). يضاف ذلك إلى تغذية الزجاج. </li><li style=";text-align:right;direction:rtl"> توصيل وحدة التغذية إلى المياه الدافئة ، سترة (37 درجة مئوية) ، وتوضع على قفص للسماح بالوصول البعوض على سطح الغشاء. </li><li style=";text-align:right;direction:rtl"> يسمح للبعوض لتغذية لمدة 30 دقيقة ~. </li></ol></li></ol> اليوم 3 : <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> والإناث تضع البيض بعد يومين تتغذى على الدم. يتم وضع ورقة صغيرة ملفوفة في تصفية شكل مخروطي في كوب صغير يحتوي على الماء المقطر ، والتأكد من أن يحصل مرشح ورقة رطبة. يتم الاحتفاظ الدورق داخل القفص ليلة وضحاها البعوض لوضع البيض. </li></ol> اليوم 4 : <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> يتم وضع ورقة الترشيح التي تحتوي على بيض البعوض في علبة بلاستيكية بالماء المقطر 300 مل ~. وقليل من الطعام ويضاف إلى الدرج ويسمح ليفقس البيض إلى اليرقات خلال الايام القليلة المقبلة. </li></ol> 5 أيام -- 8 : <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> ويتم تغذية يرقات تزداد كل يوم مع اثنين من أقراص سي الغذائية ومراقبتها لكثافة والسكان. في اليوم الثامن (اليرقات البالغة من العمر 5 أيام) ، وتضعف السكان اليرقات من 1 إلى درج الصواني 10 ~ ، مع قليل من الغذاء وقرصين من C الغذائي في كل درج. (~ 30 دقيقة) </li></ol> يوم 9 -- 12 : <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> ويتم تغذية يرقات كل يوم مع جيم الغذاء في اليوم 12th (9 اليرقات يوم من العمر) ، يتم تغيير المياه مع المياه العذبة ، وإضافة المواد الغذائية (B قليل من الطعام وقرصين من C الغذاء). ويبدأ الشرانق النامية في هذه المرحلة. وتغطي شبكات مع الأدراج لتجنب هروب الكبار. (~ 80-10 دقيقة) </li></ol> يوم 13 -- 15 : يسمح للعذارى في الظهور بين الكبار ل2 التالي -- 3 أيام. يعطى الغذاء كل يوم إلى يرقات / الشرانق عن طريق إزالة بعناية لتجنب هروب الصافي من البالغين. اليوم 16 : <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> يتم جمعها من البالغين في قفص من خلال الشافطة متصلة فراغ. القفص عبارة عن زجاجة صغيرة 100 مل مع فتيلة القطن التي يتم نقعها مع السكروز 10 ٪ (تعقيمها) وبطانة منشفة ورقية على الجزء السفلي لامتصاص أي انسكابات السكر المحتملة التي قد تحدث أثناء التعامل مع القفص. (~ دقائق 30-40) </li></ol> اليوم 17 -- 21 : <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> ثم يتم الاحتفاظ من البالغين (ذكور واناث) في غرفة insectary لأيام 4-5 ، يتغذى على السكروز 10 ٪ مرة أخرى قبل أن يتم تغذيتها بالدم لبدء الدورة القادمة. يمكن استخدام نفس البعوض لوضع البيض أكثر من مرة. </li><li style=";text-align:right;direction:rtl"> ويمكن قتل البعوض الذي ليست هناك حاجة لاجراء تجارب تربية أو عن طريق وضع القفص في الفريزر. الأدراج والأقفاص المستخدمة تحتاج إلى تنظيفها وتجفيفها قبل أن يمكن استخدامها مرة أخرى. </li></ol> ملاحظة : يمكن أن يكون هناك بعض الاختلافات في طريقة تربية البعوض ومختبرات مختلفة قد تكون التقنيات المختلفة.

<ol>
	<li>Bangs, M. J., Soelarto, T., Barodji, B. P., Wicaksana, D. T., Boewono, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Colonization+of+Anopheles+maculatus+from+Central+Java,+Indonesia.">Colonization of Anopheles maculatus from Central Java, Indonesia.</a> J Am Mosq Control Assoc. 18, 359-363 (2002).</li><li>Beier, M. S., Beier, J. C., Merdan, A. A., el Sawaf, B. M., Kadder, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laboratory+rearing+techniques+and+adult+life+table+parameters+for+Anopheles+sergentii+from+Egypt.">Laboratory rearing techniques and adult life table parameters for Anopheles sergentii from Egypt.</a> J Am Mosq Control Assoc. 266, 266-270 (1987).</li><li>Benedict, M. Q., Crampton, J. M., Beard, C. B., Louis, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Care and maintenance of anopheline mosquitoes, The molecular biology of disease vectors: A methods manual. , 3-12 (1997).</li><li>Casanges, A. H., Mcgovran, E. R., Chiles, J. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rearing+of+Anopheles+quadrimaculatus+Say+and+Aedes+aegypti+(L.)+in+the+laboratory.">Rearing of Anopheles quadrimaculatus Say and Aedes aegypti (L.) in the laboratory.</a> Mosq News. 9, 112-117 (1949).</li><li>Merritt, R. W., Dadd, R. H., Walker, E. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Feeding+behavior,+natural+food+and+nutritional+relation+ships+of+larval+mosquitoes.">Feeding behavior, natural food and nutritional relation ships of larval mosquitoes.</a> Annual Rev. Entomol. 37, 349-376 (1992).</li><li>Nasirian, H., Ladonni, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Artificial+bloodfeeding+of+Anopheles+stephensi+on+a+membrane+apparatus+with+human+whole+blood.">Artificial bloodfeeding of Anopheles stephensi on a membrane apparatus with human whole blood.</a> J Am Mosq Control Assoc. 22, 54-56 (2006).</li><li>Ng'habi, K. R., John, B., Nkwengulila, G., Knols, B. G., Killeen, G. F., Ferguson, H. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+larval+crowding+on+mating+competitiveness+of+Anopheles+gambiae+mosquitoes.">Effect of larval crowding on mating competitiveness of Anopheles gambiae mosquitoes.</a> Malar J. 30 (4), 49 (2005).</li><li>Stobart, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Factors+affecting+the+control+of+body+volume+in+the+larvae+of+the+mosquitoes+Aedes+aegypti+and+Aedes+detritus.">Factors affecting the control of body volume in the larvae of the mosquitoes Aedes aegypti and Aedes detritus.</a> Edw. J. Exp. Boil. 54, 67-82 (1971).</li></ol>

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		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Food A</td>
<td>&nbsp;</td>
<td>Aquaricare</td>
<td>&nbsp;</td>
<td>Grounded fish food. A small pinch needs to be added.</td>
</tr>
<tr>
<td>Food B</td>
<td>&nbsp;</td>
<td>Purina</td>
<td>&nbsp;</td>
<td>Grounded cat food. A small pinch needs to be added.</td>
</tr>
<tr>
<td>Food C</td>
<td>&nbsp;</td>
<td>Purina</td>
<td>&nbsp;</td>
<td>Cat food. Two tablets needs to be added</td>
</tr>
<tr>
<td>RBC</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>prepared red blood cells: 40% hematocrit in human serum</td>
</tr>
<tr>
<td>RPMI</td>
<td>Medium</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>CPD</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>anticoagulant</td>
</tr>
<tr>
<td>serum</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>O+ human serum</td>
</tr>
<tr>
<td>Mosquito feeder</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>parafilm</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Anopheles gambiae</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>mosquitos</td>
</tr>
<tr>
<td>10% sucrose</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>mosquito food</td>
</tr>
<tr>
<td>insectary</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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protocol for mosquito rearing (a. gambiae)

يصف هذا البروتوكول تربية البعوض في insectary. يتم الحفاظ على غرف insectary في 28 درجة مئوية والرطوبة 80 ٪ ~ ، مع 12 ساعة. اليوم / دورة الليل. لهذا الإجراء ، ستحتاج أقفاص البعوض ، 10 ٪ محلول السكروز العقيمة ، والمناشف الورقية ، دورق ، whatman رقة الترشيح ، مغذيات الزجاج ، ومصل دم الإنسان وحمام المياه ، parafilm ، الماء المقطر ، والصواني البلاستيكية النظيفة والغذاء البعوض (وصفه أدناه) ، والبعوض صافي لتغطية الصواني ، والفراغ ، وغرفة لجمع جمع الكبار.

Watch this Scientific Journal Video about Protocol for Mosquito Rearing A. gambiae at JoVE.com

Protocol for Mosquito Rearing A. gambiae

هذا الفيديو يوضح التقنيات العامة المستخدمة لالأنوفيلة الغامبية الخلفي في المختبر. وأثبتت الأساليب لرعاية البعوض المختبرات من خلال جميع مراحل دورة حياة الكائن الحي من اليرقات إلى الشرانق لتغذية الدم الكبار.

بروتوكول للتربية البعوض (A. الغامبية)

This protocol describes mosquito rearing in the insectary. The insectary rooms are maintained at 28&deg;C and ~80% humidity, with a 12 hr. day/night ...

Research

JoVE Journal

Biology

Protocol for Mosquito Rearing (A. gambiae)

23.0K Views.  Johns Hopkins University. هذا الفيديو يوضح التقنيات العامة المستخدمة لالأنوفيلة الغامبية الخلفي في المختبر. وأثبتت الأساليب لرعاية البعوض المختبرات من خلال جميع مراحل دورة حياة الكائن الحي من اليرقات إلى الشرانق لتغذية الدم الكبار.

Video: Protocol for Mosquito Rearing A. gambiae

Protocol for RNAi Assays in Adult Mosquitoes (A. gambiae)

Reverse genetic approaches have proven extremely useful for determining  which genes underly resistance to vector pathogens in mosquitoes.  This video protocol illustrates a method used by the Dimopoulos lab to inject dsRNA into Anopheles gambiae mosquitoes, which harbor the malaria parasite.  The technique manipulating the injection setup and injecting dsRNA into the thorax is illustrated.

Protocol for RNAi Assays in Adult Mosquitoes A. gambiae

Reverse genetic approaches have proven extremely useful for determining which genes underly resistance to vector pathogens in mosquitoes. This video protocol illustrates a method used by the Dimopoulos lab to inject dsRNA into Anopheles gambiae mosquitoes, which harbor the malaria parasite. The technique manipulating the injection setup and injecting dsRNA into the thorax is illustrated.

بروتوكول للفحوصات رني في البعوض الكبار (A. الغامبية)

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

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

In this interview, George Dimopoulos focuses on the physiological mechanisms used by mosquitoes to combat Plasmodium falciparum and dengue virus infections.   Explanation is given for how key refractory genes, those genes conferring resistance to vector pathogens, are identified in the mosquito and how this knowledge can be used to generate transgenic mosquitoes that are unable to carry the malaria parasite or dengue virus.

In this interview, George Dimopoulos focuses on the physiological mechanisms used by mosquitoes to combat Plasmodium falciparum and dengue virus infections. Explanation is given for how key refractory genes, those genes conferring resistance to vector pathogens, are identified in the mosquito and how this knowledge can be used to generate transgenic mosquitoes that are unable to carry the malaria parasite or dengue virus.

بناء عالم أفضل البعوض : تحديد الجينات تمكين الملاريا وحمى الضنك حمى المقاومة في الغامبية ألف وألف البعوض بعوض

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

A Protocol for the Production of KLRG1 Tetramer

Killer cell lectin-like receptor G1 (KLRG1) is a type II transmembrane glycoprotein inhibitory receptor belonging to the C type lectin-like superfamily. KLRG1 exists both as a monomer and as a disulfide-linked homodimer. This well-conserved receptor is found on the most mature and recently activated NK cells as well as on a subset of effector/memory T cells.
Using KLRG1 tetramer as well as other methods, E-, N-, and R-cadherins were identified as KLRG1 ligands. These Ca2+-dependent cell-cell adhesion molecules comprises of an extracellular domain containing five cadherin repeats responsible for cell-cell interactions, a transmembrane domain and a cytoplasmic domain that is linked to the actin cytoskeleton. 
Generation of the KLRG1 tetramer was essential to the identification of the KLRG1 ligands. KLRG1 tetramer is also a unique tool to elucidate the roles cadherin and KLRG1 play in regulating the immune response and tissue integrity.

This protocol describes the production of KLRG1 tetramer, which is a powerful tool for the analysis of KLRG1 ligands.

بروتوكول لإنتاج Tetramer KLRG1

Killer cell lectin-like receptor G1 (KLRG1) is a type II transmembrane glycoprotein inhibitory receptor belonging to the C type lectin-like superfamily. ...

An Optimized Protocol for Rearing Fopius arisanus, a Parasitoid of Tephritid Fruit Flies

Fopius arisanus (Sonan) is an important parasitoid of Tephritid fruit flies for at least two reasons. First, it is the one of only three opiine parasitoids known to infect the host during the egg stage1. Second, it has a wide range of potential fruit fly hosts. Perhaps due to its life history, F. arisanus has been a successfully used for biological control of fruit flies in multiple tropical regions2-4. One impediment to the wide use of F. arisanus for fruit fly control is that it is difficult to establish a stable laboratory colony5-9. Despite this difficulty, in the 1990s USDA researchers developed a reliable method to maintain laboratory populations of F. arisanus10-12. There is significant interest in F. arisanus biology13,14, especially regarding its ability to colonize a wide variety of Tephritid hosts14-17; interest is especially driven by the alarming spread of Bactrocera fruit fly pests to new continents in the last decade18. Further research on F. arisanus and additional deployments of this species as a biological control agent will benefit from optimizations and improvements of rearing methods. In this protocol and associated video article we describe an optimized method for rearing F. arisanus based on a previously described approach12. The method we describe here allows rearing of F. arisanus in a small scale without the use of fruit, using materials available in tropical regions around the world and with relatively low manual labor requirements.

An Optimized Protocol for Rearing Fopius arisanus, a Parasitoid of Tephritid Fruit Flies

Fopius arisanus is an egg-larval parasitoid of Tephritid fruit flies that is successfully used in biological control of these important tropical pests. We describe here an optimized protocol for rearing F. arisanus on a small scale using readily available materials.

بروتوكول الأمثل لتربية Fopius arisanus ، وهو الطفيل من الفاكهة Tephritid الذباب

Fopius arisanus (Sonan) is an important parasitoid of Tephritid fruit flies for at least two reasons. First, it is the one of only three opiine ...

SIVQ-LCM Protocol for the ArcturusXT Instrument

SIVQ-LCM is a new methodology that automates and streamlines the more traditional, user-dependent laser dissection process.&#160;It aims to create an advanced, rapidly customizable laser dissection platform technology. In this report, we describe the integration of the image analysis software Spatially Invariant Vector Quantization (SIVQ) onto the ArcturusXT instrument. The ArcturusXT system contains both an infrared (IR) and ultraviolet (UV) laser, allowing for specific cell or large area dissections. The principal goal is to improve the speed, accuracy, and reproducibility of the laser dissection to increase sample throughput. This novel approach facilitates microdissection of both animal and human tissues in research and clinical workflows.

SIVQ-LCM is an innovative approach that harnesses a computer algorithm, Spatially Invariant Vector Quantization (SIVQ), to drive the laser capture microdissection (LCM) process. The SIVQ-LCM workflow greatly improves the speed and accuracy of microdissection, with applications in both the research and clinical settings.

بروتوكول SIVQ-LCM لالصك ArcturusXT

SIVQ-LCM is a new methodology that automates and streamlines the more traditional, user-dependent laser dissection process.&#160;It aims to create an ...

An Experimental and Bioinformatics Protocol for RNA-seq Analyses of Photoperiodic Diapause in the Asian Tiger Mosquito, Aedes albopictus

Photoperiodic diapause is an important adaptation that allows individuals to escape harsh seasonal environments via a series of physiological changes, most notably developmental arrest and reduced metabolism. Global gene expression profiling via RNA-Seq can provide important insights into the transcriptional mechanisms of photoperiodic diapause. The Asian tiger mosquito, Aedes albopictus, is an outstanding organism for studying the transcriptional bases of diapause due to its ease of rearing, easily induced diapause, and the genomic resources available. This manuscript presents a general experimental workflow for identifying diapause-induced transcriptional differences in A. albopictus. Rearing techniques, conditions necessary to induce diapause and non-diapause development, methods to estimate percent diapause in a population, and RNA extraction and integrity assessment for mosquitoes are documented. A workflow to process RNA-Seq data from Illumina sequencers culminates in a list of differentially expressed genes. The representative results demonstrate that this protocol can be used to effectively identify genes differentially regulated at the transcriptional level in A. albopictus due to photoperiodic differences. With modest adjustments, this workflow can be readily adapted to study the transcriptional bases of diapause or other important life history traits in other mosquitoes.

An Experimental and Bioinformatics Protocol for RNA-seq Analyses of Photoperiodic Diapause in the Asian Tiger Mosquito, Aedes albopictus

RNA-Seq analyses are becoming increasingly important for identifying the molecular underpinnings of adaptive traits in non-model organisms. Here, a protocol to identify differentially expressed genes between diapause and non-diapause Aedes albopictus mosquitoes is described, from mosquito rearing, to RNA sequencing and bioinformatics analyses of RNA-Seq data.

والتجريبية وبروتوكول المعلوماتية الحيوية لتحليل-RNA وما يليها من دور التعرض للضوء الكمون في النمر الآسيوي البعوض،<em&gt; بحمى الضنك</em

Photoperiodic diapause is an important adaptation that allows individuals to escape harsh seasonal environments via a series of physiological changes, ...

RNAi Trigger Delivery into Anopheles gambiae Pupae

RNA interference (RNAi), a naturally occurring phenomenon in eukaryotic organisms, is an extremely valuable tool that can be utilized in the laboratory for functional genomic studies. The ability to knockdown individual genes selectively via this reverse genetic technique has allowed many researchers to rapidly uncover the biological roles of numerous genes within many organisms, by evaluation of loss-of-function phenotypes. In the major human malaria vector Anopheles gambiae, the predominant method used to reduce the function of targeted genes involves injection of double-stranded (dsRNA) into the hemocoel of the adult mosquito. While this method has been successful, gene knockdown in adults excludes the functional assessment of genes that are expressed and potentially play roles during pre-adult stages, as well as genes that are expressed in limited numbers of cells in adult mosquitoes. We describe a method for the injection of Serine Protease Inhibitor 2 (SRPN2) dsRNA during the early pupal stage and validate SRPN2 protein knockdown by observing decreased target protein levels and the formation of melanotic pseudo-tumors in SRPN2 knockdown adult mosquitoes. This evident phenotype has been described previously for adult stage knockdown of SRPN2 function, and we have recapitulated this adult phenotype by SRPN2 knockdown initiated during pupal development. When used in conjunction with a dye-labeled dsRNA solution, this technique enables easy visualization by simple light microscopy of injection quality and distribution of dsRNA in the hemocoel.

RNAi Trigger Delivery into Anopheles gambiae Pupae

RNA interference (RNAi) is an extremely valuable tool for uncovering gene function. However, the ability to target genes using RNAi during pre-adult stages is limited in the major human malaria vector Anopheles gambiae. We describe an RNAi protocol to reduce gene function via direct injection during pupal development.

رني الزناد تسليم إلى<em&gt; بعوض الملاريا</em&gt; شرانق

RNA interference (RNAi), a naturally occurring phenomenon in eukaryotic organisms, is an extremely valuable tool that can be utilized in the laboratory ...

IR-TEx: An Open Source Data Integration Tool for Big Data Transcriptomics Designed for the Malaria Vector Anopheles gambiae

IR-TEx is an application written in Shiny (an R package) that allows exploration of the expression of (as well as assigning functions to) transcripts whose expression is associated with insecticide resistance phenotypes in Anopheles gambiae mosquitoes. The application can be used online or downloaded and used locally by anyone. The local application can be modified to add new insecticide resistance datasets generated from multiple -omics platforms. This guide demonstrates how to add new datasets and handle missing data. Furthermore, IR-TEx can be completely and easily recoded to use-omics datasets from any experimental data, making it a valuable resource to many researchers. The protocol illustrates the utility of IR-TEx in identifying new insecticide resistance candidates using the the microsomal glutathione transferase, GSTMS1, as an example. This transcript is upregulated in multiple pyrethroid resistant populations from C&#244;te D&#39;Ivoire and Burkina Faso. The identification of co-correlated transcripts provides further insight into the putative roles of this gene.

IR-TEx: An Open Source Data Integration Tool for Big Data Transcriptomics Designed for the Malaria Vector Anopheles gambiae

IR-TEx explores insecticide resistance-related transcriptional profiles in the species Anopheles gambiae. Provided here are full instructions for using the application, modifications for exploring multiple transcriptomic datasets, and using the framework to build an interactive database for collections of transcriptomic data from any organism, generated in any platform.

IR-تكس: أداه تكامل البيانات المفتوحة المصدر لنسخ البيانات الكبيرة المصممة لمكافحه ناقلات الملاريا

IR-TEx is an application written in Shiny (an R package) that allows exploration of the expression of (as well as assigning functions to) transcripts ...

A Step-by-Step Guide to Mosquito Electroantennography

Female mosquitoes are the deadliest animals on earth, claiming the lives of more than 1 million people every year due to pathogens they transmit when acquiring a blood-meal. To locate a host to feed on, mosquitoes rely on a wide range of sensory cues, including visual, mechanical, thermal, and olfactory. The study details a technique, electroantennography (EAG), that allows researchers to assess whether the mosquitoes can detect individual chemicals and blends of chemicals in a concentration-dependent manner. When coupled with gas-chromatography (GC-EAG), this technique allows to expose the antennae to a full headspace/complex mixture and determines which chemicals present in the sample of interest, the mosquito can detect. This is applicable to host body odors as well as plant floral bouquets or other ecologically relevant odors (e.g., oviposition sites odorants). Here, we described a protocol that permits long durations of preparation responsiveness time and is applicable to both female and male mosquitoes from multiple genera, including Aedes, Culex, Anopheles, and Toxorhynchites mosquitoes. As olfaction plays a major part in mosquito-host interactions and mosquito biology in general, EAGs and GC-EAG can reveal compounds of interest for the development of new disease vector control strategies (e.g., baits). Complemented with behavioral assays, the valence (e.g., attractant, repellent) of each chemical can be determined.

The present article details a step-by-step protocol for successful and low noise electroantennograms in several genera of mosquitoes, including both females and males.

دليل خطوة بخطوة لتخطيط كهربية البعوض

Female mosquitoes are the deadliest animals on earth, claiming the lives of more than 1 million people every year due to pathogens they transmit when ...

A Magnetic-Bead-Based Mosquito DNA Extraction Protocol for Next-Generation Sequencing

A recently published DNA extraction protocol using magnetic beads and an automated DNA extraction instrument suggested that it is possible to extract high quality and quantity DNA from a well-preserved individual mosquito sufficient for downstream whole genome sequencing. However, reliance on an expensive automated DNA extraction instrument can be prohibitive for many laboratories. Here, the study provides a budget-friendly magnetic-bead-based DNA extraction protocol, which is suitable for low to medium throughput. The protocol described here was successfully tested using individual Aedes aegypti mosquito samples. The reduced costs associated with high quality DNA extraction will increase the application of high throughput sequencing to resource limited labs and studies.

Described here is a DNA extraction protocol using magnetic beads to produce high quality DNA extractions from mosquitoes. These extractions are suitable for a downstream next-generation sequencing approach.

بروتوكول استخراج الحمض النووي للبعوض القائم على الخرز المغناطيسي لتسلسل الجيل التالي

A recently published DNA extraction protocol using magnetic beads and an automated DNA extraction instrument suggested that it is possible to extract high ...