Trial, Error, and Learning
The saying, "There cannot be success without failure" is true for scientific research. In many cases, you will do an experiment multiple times and still get the same unsuccessful results. When this happens, you cannot take it as a personal defeat, but rather as a time for reflection and identify any changes to variables in order to pick yourself back up and try again.
Each time you repeat an experiment you gain more knowledge and awareness on the procedure and technical skills required for each task. Even though you may get unfavorable results, you learn more and more about what could have gone wrong and take proper action to change it. I am all too familiar with the trial and error of research.
Throughout the summer, I have been working on microinjection experiments to identify mutations in pigment genes in zebrafish. This experiment is used as a baseline control to ensure the microinjections are working properly. However, I have spent a majority of my summer in lab trying to get sufficient mutations in order to move on to our target gene. The first thing I did was tell myself not to get discouraged because it is a learning process and even the most experienced researchers still get insufficient results. The next thing I do is go through the experimental process and check if there was anything I did differently this time compared to previous experiments. If there is nothing different, then I see what is variable in the experiment that could yield these results. In my case, there are a few variables that could yield insufficient results;
old/expired Cas9 aliquots
old/expired gRNA
timing of injections into the embryo (single-cell vs. multi-cell)
the amount of reagents being injected into the embryo
All of these factors can influence the amount of mutations seen in any given round of microinjections.
What I have learned most from lab and the trial and error of experiments is to have patience, trust in yourself, have resiliency, and rid your mind of imposter syndrome thoughts. There are some difficult and technical processes and experiments required, but you will only get better with practice and having the belief in yourself that you are capable of carrying them out. If you continuously think you are not good enough and that you don't belong, it will start to show in your work and negatively affect your performance and knowledge in the lab.
"Success is not final, failure is not fatal: it is the courage to continue that counts"
~Winston S. Churchill
