I previously published a blog post called Going through the Alphabet, where I talked about the trials and tribulations that I faced during the process of nailing down my project.
Since then, I’ve learned that the same persistence is required for the data collection stage.
My project has three major components: 1) testing Mongolian Oaks for bacterial leaf scorch by extracting DNA and using a polymerase chain reaction (PCR), 2) determining moisture content by measuring fresh and dry weight, and 3) determining the percentage of foliar Nitrogen present.
The first and third components in particular require several steps and depend on a variety of factors. Extracting DNA and using PCR involves a tremendous amount of pipetting small volumes of liquids. To visualize your results, you must run a gel electrophoresis, which can be finicky at times. In addition to the hours of benchwork required by these three steps, you must account for the time required for instruments to run. Once you have amplified your DNA using PCR, you must put your samples in the thermocycler for about two hours. Gels take at least an hour to set up and run. Use of instruments is also limited by availability and space within the instrument. I have fifty samples, which translates to three rounds of DNA extraction, four rounds of PCR, and three gels. The number of steps within each of these three stages, coupled with the repetitions required by the number of samples I am testing and the mistakes that I have made with pipetting, results in a very exhausting and time-consuming process.
Determining foliar N levels has three stages as well. First, leaves must be ground to dust using a coffee grinder and a mortar and pestle. Then, you wrap 5 mg of each sample into a tiny tin boat. Finally, you run these tinned samples through an Elementar, which burns them at an extremely high temperature. Technically, this analysis does not require rounds like my genetic work does. However, due to mistakes that I have made and technical issues with the Elementar, my samples have had to be re-tinned twice.
My project has been further hindered by mistakes that I have made. Although several of my first attempts did not succeed due to mistakes I made with pipetting and with tinning, and other attempts have gone awry due to malfunctions with instruments, I have gained from my experiences and learned a great deal from my mistakes. I think the most important realization I have made during my time here is that research requires more than just persistence, attention to detail, and a willingness to ask questions. It requires optimism - the ability to remain positive and keep trying when things fail or when you make mistakes.
Even though research can be incredibly painful at times, it definitely pays off. Nothing is more satisfying than looking at the raw data you collected and knowing that it was a product of your hard work and determination, as well as the support of others. Even though roadblocks and mistakes are bound to happen, if you stay positive and keep trying, success in research will eventually happen too.