Training Complete

For the past few weeks, Dr. Hinton has trained me and my fellow summer lab members on various procedures and experimental techniques that are commonly used in lab. We have learned how to properly use the tissue culture room, which is a sterile room where we are able to culture various cell lines. There, we are able to treat our cells with different plasmids, giving them different expressions, and under treatment, we can see how these different expressions react under treatments. Along with that we have learned how to make more plasmids when we are running low. This process can be extensive, especially when they are multiple plasmids that need to be replenished. We used E. Coli bacteria to help culture the plasmids. Once the bacteria was transformed with corresponding plasmids, we broke up the cytoskeleton and other extracellular membrane proteins  to isolate the plasmids for further experiments. We have also learned how to do Western blots. Western blots help detect certain proteins with the help of antibodies that are specific to the proteins of interest. Western blots are done by first running a 2D gel with an electric current. After that, the gel is placed into a machine to transfer the data onto a membrane which allows for ease with antibody tagging. Training has gone very well so far, and I cannot wait to start my independent project.

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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