Blog 11: Mentorship 10 Hours
1. Biochem Lab in Cal State Fullerton
2. Mike Suseoff
3. 28 Hours
4. (4 Hours): The graduate that I am shadowing, Mike explained the lab we would be taking part in and the procedures we would have to do during the lab and why exactly we had to do them and any other questions I had. After this we started the lab, Mike had already made a few preparations before I arrived. In short, we grew cells. Mike taught me and let me use some of the equipment that I have never had experience using before like, fixed volume pipettes, a low speed centrifuge, and a spectrometer. There was a lot of waiting for the different machines we had to use for the lab, we cleaned and sanitized everything and refilled some of the chemicals.
(4 Hours): Today, we harvested the cells we grew! We started by taking the media and cells out of the low speed centrifuge then we put them into four containers and balanced them on a scale so that each two weighed exactly the same. Then after we set them in a high speed centrifuge so that the cells and the media would seperate, the cells would become a pellet at the bottom of the container. We took them out of the high speed centrifuge and then poured out the media so we would be left with only the cells, then we scraped the cells out and put them into smaller covered containers and figured out the weight of the cells we harvested, after we put them on ice. And we continued with the process of purification..
(2 hours): I came in later than normally today so there was a lot of down time while the protein/buffer was in the centrifuge, we did this because we wanted to purify the solution more to get more protein than buffer for this we used a kind of filter to take out the sodium in the solution. This raised the concentration of our protein in the solution and lowered the concentration of the sodium. While waiting for this we started a procedure called the Bradford Assay which is used to measure the concentration of protein in a solution. We used this on the crude we extracted from our protein, it showed how much other proteins were in the crude and it came out to be around 43%.
2. Mike Suseoff
3. 28 Hours
4. (4 Hours): The graduate that I am shadowing, Mike explained the lab we would be taking part in and the procedures we would have to do during the lab and why exactly we had to do them and any other questions I had. After this we started the lab, Mike had already made a few preparations before I arrived. In short, we grew cells. Mike taught me and let me use some of the equipment that I have never had experience using before like, fixed volume pipettes, a low speed centrifuge, and a spectrometer. There was a lot of waiting for the different machines we had to use for the lab, we cleaned and sanitized everything and refilled some of the chemicals.
(4 Hours): Today, we harvested the cells we grew! We started by taking the media and cells out of the low speed centrifuge then we put them into four containers and balanced them on a scale so that each two weighed exactly the same. Then after we set them in a high speed centrifuge so that the cells and the media would seperate, the cells would become a pellet at the bottom of the container. We took them out of the high speed centrifuge and then poured out the media so we would be left with only the cells, then we scraped the cells out and put them into smaller covered containers and figured out the weight of the cells we harvested, after we put them on ice. And we continued with the process of purification..
(2 hours): I came in later than normally today so there was a lot of down time while the protein/buffer was in the centrifuge, we did this because we wanted to purify the solution more to get more protein than buffer for this we used a kind of filter to take out the sodium in the solution. This raised the concentration of our protein in the solution and lowered the concentration of the sodium. While waiting for this we started a procedure called the Bradford Assay which is used to measure the concentration of protein in a solution. We used this on the crude we extracted from our protein, it showed how much other proteins were in the crude and it came out to be around 43%.
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