When I first saw my research project for the summer and what I would need to do to accomplish it, I thought my tasks were pretty straightforward and I would be done in two weeks with clear results. Well, turns out I was somewhat naïve and uninformed about this whole doing science because that didn’t turn out to be the case at all. I was unaware of unexpected outcomes. We finished the sixth week now and I just hope that I’ll be able to get my results by the poster presentation session which is in two weeks.
My project involves mutation of a particular gene using PCR and running Western Blots on human cells after transfecting them with these mutated genes as I mentioned multiple times in my previous blogs. I need to go through this process multiple times for different regions targeted for mutations and compare the results. So far, I have been able to get my mutant genes but it took 5 weeks for me to do it which would normally take one if everything went smoothly. Of course, nothing went smoothly. I encountered problems and made mistakes at every step of the way. Sometimes, PCR didn’t work, sometimes transformation and sometimes the transformed plasmids didn’t have the desired plasmids. I had to optimize the method by adjusting the temperature, tweaking protocols and adding some different chemicals to make the experiments work. Eventually, I was able to get the mutations I wanted, and the feeling of overcoming problems and reaching my goal was quite pleasing.
Now that I finally have my mutations, I will proceed to the next steps which should take about four days –again— if everything goes smoothly. I hope that I can troubleshoot my way into the results before two weeks. Although the problems I had to cope with and the mistakes I made were occasionally frustrating, they taught me one valuable lesson about research as Thomas Edison said: “Our greatest weakness lies in giving up. The most certain way to succeed is always to try just one more time.”
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