Welcome to the first "real" post where I tell you guys about what is going on in the lab! First things first, I had to complete a pretty rigorous course on biosafety. Who would have thought that you should not eat or drink in a lab full of pathogens and caustic chemicals? In all seriousness though, I was finally able to learn the difference between all the different levels of biosafety. These levels range from one to four; one being practically harmless and difficult to contract and four being lethal and highly infectious. For reference, the lab that I am working in is BSL-2, as their are some infectious agents which are not particularly dangerous. Each level also requires different levels of protective measures. For BSL-2, I need to wear gloves, a lab coat, and sometimes eye and mouth protection, depending on what I am working with. For now though, I have not really entered that level as I am just working with E. Coli.
On to my actual experiment, I have been attempting to transform two different types of E.coli, DH5α
and BL21. Transforming a bacteria is not what you think it is (though changing how it looks would be pretty neat). All it is is just making that bacteria absorb DNA and express it, in my case a plasmid coding for pfu DNA polymerase, another type thermostable DNA polymerase, and ampicillin resistance. By giving these bacteria a resistance to ampicillin, then they can survive in an LB agar containing ampicillin, which weeds out the bacteria that did not transform. After one failed attempt though, I managed to get colonies to grow with the DH5α E. Coli., but not the BL21 unfortunately.
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| If you look closely, you can see little, white dots all over the DH5α plate. |
Next week, I will be trying to extract the pfu from the DH5α and purify it as well as figuring out why the BL21 would not grow. Stay tuned for more updates!
I remember plasmids from AP Bio; it was always an intriguing subject. Great work successfully growing a colony. The one that didn't grow might ultimately end up being more important if you determine the reason for its failure.
ReplyDeleteDylan: This is a great opportunity for you to do real science. I will be following your SRP as an individual who is fascinated by molecular biology. The implications of success in this experience are very significant. Good luck.
ReplyDeleteFascinating! I bet that the lesson on safety measures was quite interesting. Do you ever feel nervous entering the lab setting? What are the precautions for working with E Coli specifically?
ReplyDeleteThe E. Coli that I am working with is relatively harmless. I would say the biggest precaution is don't ingest it, which I really doubt I will ever do.
DeleteWow, it sounds like your project is going great! Do you know why the first transformation attempt failed?
ReplyDeleteNot entirely sure, it could have just been because the cells I was using were not competent or I may have killed them by heat shocking them for too long.
DeleteAli told me you were in Chicago (which I didn't know). Sounds super interesting so huzzah! Don't catch anything weird.
ReplyDeleteHi Dylan! Your project sounds really hands-on -- how exciting! Over the course of the next two months, do you think that you will need to use that extra protection or will you just be working with E. coli the entire time? I'm very eager to hear more about your project as time goes on.
ReplyDeleteI am probably going to be working with E. Coli most of the time. The lab is BSL-2 mostly because they also work with viruses like Polio.
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