Ruben Galindo, Post 1

June 16

            It has been about two weeks since I joined the lab, and one day I came to lab and found a large puddle of brown mystery liquid under my chair that gave off a pungent smell. I looked for the source but failed to find a tipped over bottle or leaky pipe. When my mentor arrived, he was also unable to determine the source of the liquid or its composition. After asking around, we found out that it likely came from the water pipes in the ceiling that were being worked on the previous day. So based on this assumption, we decided to just mop up the mess. This makes me pity who ever uses the eye wash station because the leaky pipes that contain the rusty water probably lead to the eye wash station. So whoever gets chemicals in their eyes and uses the eye wash, might get rusty water in their eyes.

            After the spill was cleaned up, I practiced using the emulsiflux. I was shown how to use it to lyse cells the previous day, but I was still unfamiliar with it. It is a relatively loud machine that uses a high pressure force to lyse cells. 

July 1

            In order to express my target proteins in HEK cells using the Bac-toBac Bacilovirus expression system, a lot of media is needed. Every two days, 150ml of SF9 insect cells need to be passed. These cells are needed in order to create the bacilovirus that will be used to infect HEK cells. Therefore media is needed in order to sustain these insect cells until they are needed for viral production and amplification. In order to make room for new media, and prevent confusion, expired cell culture media was thrown out. As it turns out, someone who left the lab a couple years ago had a large amount of media that expired 2 years ago. So my mentor, another postdoc in the lab, and I poured out about 1000 dollars worth of expired media. Getting to pour such a large amount of money down the drain was a new experience that was quite enjoyable. While I was not physically burning monetary bills like in certain movies, it felt like I was.

July 2

            Once my target proteins have been expressed in HEK cells, I will need to purify them for crystallization. At this time, I am not at the step to express the proteins, so I practiced the purification method with a prepared protein sample. Following the published protocol of TRP protein, one of the steps involved using size exclusion chromatography. So after being shown how to use the program and column, I got to try purifying a few samples. A few times the pressure spiked causing the machine to error, but other than this one issue, it was fairly simple.