After a nice restful weekend, I was ready for day five. Today I worked with Dr. Todd at CA. She had some of the proteins I was working on with Dr. Bermek. With Dr. Bermek I had been working with the protein UL8. Dr. Todd had different variants of the protein like UL30. We ran a protein gel with samples sent over from UNC to check them and see if they were clean samples. With Dr. Todd watching, I set up the Tetra gel box and gel apparatus ALL BY MYSELF (I was very proud to put my new skills to use). The only thing different was this time is instead of putting a plastic dam parallel to the gel, I put another gel opposite to the first gel because we needed two to fit all the samples. Once the gels were out of the refrigerator, I opened them, tore off the plastic on the bottom, removed the comb, put the gel very tight against the gasket, aligned the red and black stickers to the red and black on the gel box, poured the buffer in the between the gels, checked if it was leaking, poured the rest of the buffer into the box, and then just like that I had set up my first gel box all by myself. Once I finished my minor accomplishment, I prepared the samples. I added 4uL of 10x loading dye to each tube. This took quite a while because there were 16 samples. Then, I added 12 uL of each sample to the each tube that already had loading dye in it. After that, I put the samples in a hot water bath at 95 degrees Celsius for five minutes. Once the samples were nice and toasty, it was time to load them in the gels. Loading gels is very tricky, so I was quite nervous for this step. I didn’t want to mess anything up that would contaminate other wells because I didn’t want to re-prepare all 16 samples or cause any extra steps for Dr. Todd. Since Dr. Todd and Dr. Bermek’s gel setup was a little different, Dr. Todd showed me how to load by demonstrating on the first two lanes. I did much better loading the gel this time since I had a little bit more practice and the pipette tips were much narrower making the placement easier. I didn’t poke the gel, and each sample went into the correct lane. Just like preparing the samples, this process was very time consuming and intricate. Once I loaded all the lanes, I turned on the electric current and let the gel run for 50 minutes.
In the meantime, I helped Dr. Todd with some lab work that needed to be prepared for next year’s Biotech class. I helped organize, hole punch, and sort a list of all the primers that had already been designed with all the information about them. Then, I labeled tube after tube with the primer name and whether it was a forward or reverse primer. Each primer needed to have duplicates, so there were a lot of tubes. Once I had finished labeling the tubes, I pipetted 30 uL from the stock primers into the new labeled tube. This process was very tedious because there were a lot of caps to unscrew and a lot of pipette tip changes, but I was happy to be able to help Dr. Todd out. Once, I had finished, Dr. Todd gave me the good news that the protein gel worked out. We had no idea what the samples were like, so she was excited that they showed up on the gel. I was excited because I completed the whole process of running a gel by myself. I prepared the samples, prepared the gel box, loaded the samples, and ran the gel. It was a day full of firsts and new accomplishments.
Today was less hectic than the others days of the WEP because there were not four or five experiments going on like at UNC. It was nice to have a slower day, but I’m excited to return to the UNC Lineberger Cancer Center tomorrow for an action packed day full of new lessons and experiences.
All the samples I prepared to load into the protein gel.
The finished stained protein gel.
The Tetra gel box I set up all by myself with gels on both sides.
The gel loaded with samples.
All the tubes I labeled and added 30 uL of primers too.