Over the past 6 weeks, I’ve learned a lot. I’ve learned how to pipette the optimal mix of reagents to amplify DNA for PCR. I’ve learned how to clone DNA with a plasmid vector. I’ve learned how to perform ventricle resection on zebrafish. And most importantly, I’ve gained a better idea of what research really is.
Research definitely takes time. Whether it was waiting a day for my bacteria inoculation or 6 hours for my RNA transcription, research actually involves lots of just waiting time. Thankfully, I was able to turn the waiting time into productive work by performing other steps or reading papers. Research also involves a lot of repetition. I can’t even count how many times I ran a PCR and gel this summer. It involves lots of repetition in the sense that repeating procedures makes you better at it. But also, troubleshooting. Sometimes, a step may not work and you may be stuck on one part for a long time, trying to figure out what’s wrong. This involves repeating the step many times and changing one thing at a time, trying all possibilities to get it perfect. I actually enjoyed this troubleshooting process because it was like solving a puzzle to me.
The main part of my project was to create 4 RNA probes for my genes of interest to perform in situ hybridization. This may sound simple to do, just create four strands of complementary RNA. Easy, right? The reality was far from this expected simplicity. In fact, it took over 6 weeks to create three probes (one gene refuses to be amplified and after tons of troubleshooting will still not work) that I just finished this Tuesday. This is really exciting for me because I can finally run all my in situs with my probes. This week will be the week of results where I can see where in whole 3 days post fertilization embryos, 6 week zebrafish hearts, uninjured zebrafish adult hearts, and injured zebrafish adult hearts the genes are being expressed. I cant wait to complete these final stages for my in situs.
In the journey to create RNA probes, I got to learn and experience a multitude of diverse procedures and techniques. Now, I feel much more comfortable performing these protocols and feel that I have a comprehensive overview of the common protocols related to zebrafish. Just the journey and learning everything both conceptually and mechanically have been so rewarding. At this point, results don’t even matter. This experience has been much more valuable (of course, it wouldn’t hurt to get some results).
Slowly by surely, I’ve been putting all the little puzzle pieces together. The past 6 weeks have gone by so quickly, I can hardly believe that the program is drawing near its end. It’s been such an enriching and thrilling experience working in the Poss Lab and I can’t wait to see the final results of my project!
