I never thought I would say this, but thank goodness one of my experiments didn’t work out! The experiment in question was the one on RNAi, during which I would have to move (and count) 450 worms a day for three days. I put the rrf/tm homozygous line that I bred on the special RNAi plates, waiting for the 2nd generation whose genes should definitely be turned off (normally the 1st generation is not on the plates for enough of their lifespan to completely do so) to be in the 4th stage of life, but the 1st generation looked really, really sick and didn’t even lay eggs. Even though it’s not what we planned, it’s still an interesting observation. Often in science, a lack of results actually turns out to be results of their own. Because this experiment didn’t pan out, my prediction of having to spend so much time in the lab was thankfully wrong.
In other news, we performed qPCR to measure the gene expression of worms infected with 4 different pathogens (the 2nd repetition of this experiment). Unlike my first qPCR, this one’s results seem much more credible. The graph below shows the relative expression of the gene in question, nspc-14, in different lines and on different bacteria.
HB (HB101) is the wildtype bacteria that are used to grow worms under normal conditions-our control. PAO1 is P. aeruginosa, a strain of this pathogen that is harmful just to worms. SM is S. marcescens and SA is S. aureus, both bacteria that are very pathogenic not just to worms, but to humans as well. N2 and TM are our main worm lines- N2 is the untouched wildtype, and TM3504 is a line that lacks the pqn-44 gene. nspc-14, the gene that is specifically being looked at in this graph, is hypothesized to work with and be related to to the expression pqn-44, both of which are thought to influence innate immunity. Given this, it makes sense that genes responsible for an immune response are upregulated on pathogenic bacteria (PAO1, SM, SA). It also makes sense that TM always has a lower expression than N2, since it is missing the pqn-44 gene. HB N2 is not included on this graph (it should be on the very left) because something went awry with the control for baseline gene expression, so it showed expression for every other gene to be very high, making the other bars look insignificant on the scale. I am performing the infection a third time, though, so we’ll have that data next week, hopefully before I fly out on Wednesday.
I’m so sad this is my last week!!! I can’t wait to come back to my family, but I’ll miss living in Warsaw and working in this lab 😦