What a week!

So, what better way to end this work filled, satisfying week than to wake up to the sun shinig in the clear blue sky? Yes, its sunny in Paris this weekend, and you bet I'm gonna take advantage of it!

To recap, last week was not very productive on the experimental side of things, but I did do quite a bit of drawing and made significant progress in my character and story developement. This week has been both productive in the drawing and experiemntal parts of my stay in the lab. Anne, my colleague and ever patient teacher came back on Monday. Sadly, we did not recieve the oligonucleotides that we needed to perform the ligation of the GFP plasmid and the Strawberry genome. So, as I did the previous week, I spent three hours in the adjoining library, drawing and bobbing my head to tunes I've heard one hundred times. Tuesday was very much the same thing. I won't lie that I was beginning to despair at this point, and I began to look forward to three more days of blank white paper and rowdy music.

THEN THEY CAME. Yes, the oligonucleotides finally came. I don't think I ever been as happy to see a macromolecule as I was on Wednesday. I don't know if I ever will be again, but it was a very wondurous feeling. So on Wednesday we began the ligation of the GFP plasmid and the Strawberry genome. First, we had to actually cut the plasmid at th e right place with the right enzymes, and thats where the oligonucleotides came in handy. We first took the specific enzymes, Xba1 and BamH1 and mixed them into different volumes of water so that they could be of the same concentration but still be effective in doing their cutting, and then we added the oligonucleotides. That took a bunch of time, and we had to wait about an hour and a half to really let it get ready to work. After that, we needed to do a PCR, which is just making literally millions of the same copy of DNA. For that , we obviously needed the plasmid and the genome, and we added the buffer, an enzyme called Taq. Pol., which is basically DNA polymerase.We added some buffer and some water to give it a medium in which to work in, and left it overnight to work its magic.

Bright and early Thursday morning, We took our ligations and did some gel electrophoresis, just to see if the ligation itself had worked. I was a bit apprehensive due to the fact that I had a taste of failure in the first try with this experiment, but when we placed the gel under the UV light, we could see a very defined dark band at around 800 base pairs (of DNA), which meant the ligation was a success!

We proceeded to continue by transforming our newly formed plasmid into cultured bacteria. This process, I have since realised, is much easier than the one I had just come through, so I entered into it with new confidence and a bit of "swagger", as some might say. We performed a transformation of the plasmid into the bacteria. The important thing in this part was to keep everything really cold, especially the bacteria, so that when they entered the hot water bath, the shock they would recieve would really open their membranes and the chances of our plasmids being absorbed were heightened. During this process, I was again shown how to make the gels, which has become easier for me to understand, now that I more familiar with it. So, again, this process took about an hour, so after spending an hour with my fellow scientists laughing about non-scientific stuff (believe it or not, they are normal people!) like James Bond and Italian desserts, we came back up to set up to plates that we would grow cultures on. We prepared the mixture, which is very luch like the process of making gels for gel electrophoresis, but with a few chemicals added. We first added X-gal, a molecule that would make the bacteria glow blue if they didn't absorb the plasmid. Another molecule we added was IPTG. This process is actually really interesting but quite long to explain. On our normal plasmids, there is a gene that, when in the presence of IPTG, produces a chemical that gives off the color blue. We actually inserted the Strawberry gene right in the middle of that gene, so now, that gene cannot be expressed, the colonies will now not be blue, and we will know exactly which bacteria had absorbed the right plasmid, and which we should use( well, that wasn't too long after all)! Genius, right? Oh, and we added some ampicillin so that any bacteria who hadn't at all absorbed the plasmid, which happened to have a gene resistant to ampicillin, would die. Amazing stuff, really.

So, after preparing the plates, we placed the bacteria into some yummy LB agar, so they could feed and reproduce prodigiously, and we spread them onto our plates. Done! Well, not totally. Like always, there was the "Did it work?" question in the back of my head.

Today, I rushed into the lab to see if it worked, and lo and behold, it did! little blue and white colonies dotted the plates (and there were a lot too!) . Ah science is so gratifying when it works. So for now, we have succeed in that part of the experiment. I don't have any specifics as of right now, but I know we will be doing some verification on the Dna and growth culture on the white bacteria, since they're the ones we want. But, we are that much closer to finally performing the growth cultures!

This week has also been quite prodigious in drawing. I think I finally have finished character developement, and the story has been written, but, being the person I am, I still want to spend a little more time drawing and developing it.

So, in between getting to play a part in a movie that I got to create myself with a group of people at an exibition that recreated the process of how to make a movie and provided suitable equipment for the task, seeing True Grit with my uncle, and playing B-ball with five girls who nearly showed me up and an African from Seattle, I'd say life here is going really well. Plus, its SUNNY! Looking forward to the next week.

Tired but satisfied, this is Clover signing off.