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!
