Knockdown of CTL4 in A. coluzzii

Total RNA extracted from A. coluzzii midgut infected with P. berghei c507 at 24 hpbf was used to prepare cDNA. The cDNA was used in conjunction with primers reported in (Habtewold et al., 2008 (link)) to amplify CTL4. DsRNA was then produced using the resulting PCR product and the T7 high yield transcription kit (ThermoFisher). 0.2 μg of purified dsRNA in 69 nl was injected into the thorax of A. coluzzii mosquitoes using glass capillary needles and the Nanoject II microinjector. Two to three days post injected mosquitoes were then infected with P. berghei.

Free full text: Click here

Ukegbu C.V., Christophides G.K, & Vlachou D. (2021). Identification of Three Novel Plasmodium Factors Involved in Ookinete to Oocyst Developmental Transition. Frontiers in Cellular and Infection Microbiology, 11, 634273.

Publication 2021

T7 high yield transcription kit

Capillary Cdna Dsrna Mosquitoes Needles Primers Thorax Transcription

Corresponding Organization :

Other organizations : Imperial College London, Cyprus Institute

Top 5 similar protocols

Variable analysis

independent variables

Injection of dsRNA targeting CTL4 into A. coluzzii mosquitoes

dependent variables

Infection of A. coluzzii mosquitoes with P. berghei
Potentially, effects on P. berghei development or other mosquito phenotypes

control variables

Total RNA extracted from A. coluzzii midgut infected with P. berghei c507 at 24 hpbf
Preparation of cDNA from the extracted total RNA
Use of primers reported in Habtewold et al. (2008) to amplify CTL4
Production of dsRNA using the resulting PCR product and the T7 high yield transcription kit
Injection of 0.2 μg of purified dsRNA in 69 nl into the thorax of A. coluzzii mosquitoes using glass capillary needles and the Nanoject II microinjector
Timing of 2-3 days post-injection before infecting mosquitoes with P. berghei

positive controls

None specified

negative controls

None specified

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!