Senior Project Clover Powell

So, here I am again. Its been a pretty exhausting week, and I have a whole new respecct for what it means to work in a lab and be a scientist. My colleague Anne and I have been working on basically getting the plasmids out of their respctive bacteria and purifying the actual DNA. Then, we cut out the part of the plasmid that we wanted. So, we are working with two plasmids; one has a sequence labeled PM, and the other labeled PKK. PM has a sequence that allows the bacteria that has it (a specific strain of E. Coli) to produce a protein that glows red. PKK peforms the same function, it just produces a protein that glows green. So, we've isolated the plasmids, run them through a couple runs of gel electrophoresis, just to varify the sizes of the plasmids and that they are correct. Yesterday, Anne cut the sequences that produce the glowing proteins out of the plasmids using restriction enzymes. What we are trying for is to get these sequences and put them into the same plasmid, so that when we perform the growth competition, the info is consistent and comparable. Today, we are running that solution of plasmid through another gel (that is short for gel electrophoresis) to separate the sequences from the rest of the plasmid. We will then cut out the part of the gel with the sequences and the desired plasmid and purify that. Then, we will put that into a solution and hope that the sequences and plasmid bind together.
A problem that we've run into is that, in order to correctly bind the sequence into the desired plasmid, we have to use the same restriction enzymes when we separate the sequence and plasmid. The two plasmids we are using don't use the same enzymes, so this test is just hoping that the, somehow, the sequences and the plasmid will bind together. The "real" way to do it is to attach some base pairs that correspond to a certain enzyme, have both plasmids use the same enzyme, and then most likely, the plasmid will bind with the sequences. Phew.... that's that.
From the art side, I'm still thinking of some ideas, but it seems that one is sticking, so I'll begin the storyboarding and writing process soon. I am really excited about this part of the project, so I've been drawing like mad. Hopefully, the end product will satisfy me...
And I wanted to thank Mr. Johnston. Since I know so much vocab and have done the majority of tis experiment in your class, I don't feel lost at all. The scientists here now really respect me now! Props, Mr. J.