The Forty - Seventh Day

I got up this morning at 720. I did not go for the breakfast things as the AQI this morning was in the 250's and still is.  I had yogurt and a swig of OJ for breakfast. before leaving at 800, I was not the first person there this morning which was good as this meant that I dd not have to do the "awkward stand outside of the door" and be examined by the people passing the door to the lab as they head to their labs. I started off the morning by taking a nap for a few minutes before getting to work. When we started there was a shortage of 2 ml tubes so we stole some from the fume hood bin. Faith arrived a little late after taking the wrong direction in the subway. I told her that she has dishonored her family as a native Beijinger getting lost on the subway. She laughed at that.

Yulan has some sort of new method that I think that she wants to try. We did not get to it this morning though because while we where preparing the 2 ml tubes that we got, Wendy came over and told Faith and Yulan in Chinese that it was time for coffee. They translated to me and I was very confused as I thought that it was only Faith and me that were supposed to go have coffee. It turns out that it was a lab wide event and we all crammed into Dr. Cheng's office to enjoy a cup of Columbia's second most popular export. It was good coffee. I drank it black and had almost 2 cups of it while the peeps talked. Faith got stuck in the middle of the circle and so every time someone talked she had to turn around. 

After this Dr. Cheng headed to a meeting and we went back to the lab. The PDA made yesterday turned out well and we prepared some petri dishes with it on them. I got a batch of stuff to autoclave and started it up.  I guess I will be unloading that after the lunch break. Faith showed me a bunch of the games that they like to play in class. 2048 was one of them. I told them about Conner White and getting to the 16000 tile.

We went to lunch at about 1130 and I had some tofu and a good stir fry. The coffee this morning made me rather jittery; my system is not used to that much caffeine. 

This afternoon we extracted DNA. When I came in Yulan and Faith were nowhere to be seen, so I thought that they were still taking a nap. I started helping Leo make more PDA before they showed up. It turns out that they had been on the sixth floor getting tissue samples from the transgenic tobacco. They came back down with the samples. Today Yulan decided to try another method for the DNA extraction. She put very small glass beads in with the samples and froze them in liquid Nitrogen before putting them into a shaker, a more violent shaker that is. This was supposed to break up and grind the sample. It would have worked well except the samples heated up too quickly for it to be very effective. In order to get them ground up we would have had to be pouring N2 over them while they were being shook, which would not have been feasible as the machine was one with a cover that had to be down while it ran. We did the DNA extraction using the kit again. 

I got a little dehydrated, by my terms that is, (this means that I had not drunk a liter in the past 4 hours) but drank a liter of water to rehydrate.

We finished the extraction and tested the concentration of the DNA with the nanodrop machine. The concentrations were very low. We are going to have to do PCR, a lot of it, to get them up to usable concentrations. 

After getting everything done with that, I went down and talked to Ben for a few minutes before coming back up and sitting with Yulan as she collected some samples from the dishes of fungus. I was talking to her and I found out a very important fact about what we were doing. The vector that the Agrobacteria produce does not just interfere with the expression of the gene; the vector cuts the gene out of the genome and replaces it with an antibiotic resistance.  This cleared up some questions that I had been having. After the initial culturing of the co cultivated fungus and bacteria, the individual fungus lines collected are basically selfed down for three generations (they reproduce asexually) to ensure that the gene is stable in the fungus and to isolate a single genetic line.

This made a lot more sense. However it also means that I will probably not be able to get any actual results from here until a few weeks after I finish my internship...

After that she left to go get food so I left. I came to my room and then left to go to the Sesh Wan place where I had fried rice and 4 lamb kabobs. I may have to go get more food here. I knocked on Silvio's door, but I think that he is asleep.

Good night and God bless.

getting sampler ready to extract 

co cultivation

the sun today

bottle for dave