Week 6 – Tadpole issues

Last week I diagnosed my molecular reagent issues, so this week I initiated the second frog mating. Apparently these couples aren’t into each other very much. As a result, the embryos I got aren’t as healthy as the first round I got. Many containers soon became cloudy and eggs are clearly unfertilized. These unhealthy eggs will explode in a week, affecting other healthy eggs as well. Therefore, I went back to my old tadpoles to test my hypothesis that the cause of unclear marking is the amount of protein expressed in tadpoles. Old tadpoles are bigger than what I want but are still useful if I am doing only a diagnosis experiment. I chose another mitochondria marker and reduced the amount of DNA I injected into tadpoles to see if anything has changed. The results were quite interesting. No matter how little the tag-RFP-T was expressed, it seems like it is not specific at all, while the other marker I used labeled mitochondria clearly.

Thus, my hypothesis was not supported. Regardless of the volume, the protein I used at the first time seems to be harmful to mitochondria. I decided to use another kind of  florescent protein. Unfortunately, my tadpoles aren’t healthy enough to support my further research. Therefore, the result may need to be published a bit later than I expected.