Experiments

For a full list of detailed protocols, please visit our protocols page. This page will just show an informative pipeline of the order of our experiments, making them easy to exactly replicate. See the main results of our experiments here. The specific activities that were performed by each team member are shown on a day to day basis here, where all development (successes and failures) are documented.

During our experiments we found better protocols that helped us improve our project. We also found some that we wanted to use, but couldn't due to either lack of time or resources. As you will see from our results page, the detection and testing side of our siRNAs was the main shortcoming of our project. This is because we lacked the resources for dedicated siRNA detection kits, such as the one found here. Then, we lacked the lab time to perform the aphid toxicity experiments, due to aphid generation time and needing multiple generations for reliable results. Had we had the time, we would have performed an experiment analogous to that of Rodriguez et al. in 2014⁽¹⁾.

References

(1) - Rodriguez, P., Hogenhout, S. and Bos, J. (2014). Leaf-Disc Assay Based on Transient Over-Expression in Nicotiana benthamiana to Allow Functional Screening of Candidate Effectors from Aphids. Methods in Molecular Biology:137-143.

Describe the research, experiments, and protocols you used in your iGEM project. These should be detailed enough for another team to repeat your experiments.

Please remember to put all characterization and measurement data for your parts on the corresponding Registry part pages.

• For a full list of detailed protocols, see our Protocols page
• Experiments for GFP and mCherry, in plants and in bacteria too. GUS staining assays. siRNA detection. Lack of RNAi lethality assays due to time limits. Gels and links to the results and/or gallery for the different parts in each stage.
• Images and results page will show development of the project. See the notebook page for a detailed description of activities on each day.