Protocols, Training, and First Western Blot

My first few weeks here on campus were full of learning procedures and protocols so that I could start doing experiments. The first week and a half, I began by learning how to make plasmids, which are circular strands of DNA that replicate independently. Plasmids can be used to manipulate cells to see what happens when you over- or under-express certain proteins. For example, in my lab’s most recently published paper (Banks et al 2017), we found that when MK-STYX is overexpressed in primary neurons, it changes their morphology. Plasmid preparation consists of transforming the target DNA into E. coli cells and then isolating the DNA from the bacteria after letting them multiply. I then determined the concentration of the plasmids using the Nanodrop.

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Early June Update (A Little Late)

Our experiment on milkweed pollination needed to start quickly.  Since we needed fresh blooms to quantify both longhorned milkweed beetle and general pollination, our research group needed to get to our transects out in Blandy Experimental Farm and Arboretum before any milkweed blooms.  The common milkweed starts to bloom in early June, so everything had to be ready before then.  Along with graduate student Nicki, who is running this project, another undergraduate assistant, Angelica, and I rushed to prepare all the tools and equipment we would need.  We got the lab and the greenhouse cleaned and organized, including a painstaking power-wash of the greenhouse floor.  We collected all the supplies needed, including our flag markers, binoculars, and pollination bags to protect the flowers (made by me over summer break).  When everything was collected, we woke up bright and early to head up to Blandy.

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June Update: Change of Plans

Upon returning to campus to begin my project, I found out that new data related to my project had been found while I had been away. This new data identified 398 genes whose transcription was affected (either up or down regulated) by aggregated Huntingtin’s protein (Htt). Also included was the reversal of the negative effects of Htt when a gene known as Slx5 was over-expressed. There is a lot of background information required to understand why these findings are so important.

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May 30- June 6 Establishing Controls

Stability of Relevant Molecules

For the first 2 weeks of summer research, the goal has been to lay the groundwork for the rest of the summer. The amount of time certain molecules can sit out on the lab top at room temperature before they degrade and become unusable had to be established. Then the effect of freezing and thawing an enzyme multiple times on its activity level was investigated. These things had to happen before the real research could begin. This took several days of control experiments.

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The Anticipation Builds: Preparing Data for Analysis

Hey y’all!

I’m so excited to send out my first research update! During my time in the lab thus far, I have been conducting an in-depth literature review of mentoring studies. The bulk of the literature focuses on youth characteristics that influence the mentoring relationship. These characteristics include youth environmental stress and behavioral challenges. I have only found one study that touches on mentor emotional background so I hope that I will be able to meaningfully contribute to the mentoring literature at the conclusion of my research! I will analyze mentor stress using two scales: The Risky Families Questionnaire, which measures early life and familial stress, and the Student Stress Scale, which measures previous or ongoing stress in multiple aspects of life. Combined, these two scales will allow me to not only measure mentor stress, but also will allow me to better understand the effects of different types of stressors on mentoring relationships. I will measure mentor depression using the CESDR-10 scale, and I will run analyses analyzing the relationship between mentor’s reported depression and stress levels.

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