Transmitters had been placed on three bats two days before hand. We used telemetry to try and find the bats. We only located bat 2. It was in tree 49. The myotis bats in tree 49 could be seen clearly, it was the best view i had ever had of them in the day time. We also got to hear a large colony in tree 6.
We decided to try to check the trees before the storm hit. The area was still flooded. We found two big ear bats. We got to see a tree fall and my waders filled up with water.
3/13/08 was the first day back in the field. The area was still flooded so we had to wear waders. We found two big ear bats.
So today i went into the lab and got all excited b/c i thought that i saw a colony of cells that had grown up. Later, I discovered that it was no more than a spot of dirt on the bottom of my petri dish. All the tissue culture cells were dead too. Sad day! So now we have to go back and start all over again. This entails that the DNA be re-purified, the cells be transvected, and left to grow again. This time we hope that it works.
January 26, 2008
The cells were placed in AB free medium. One of the plates just had the medium changed. One plate was treated with a plasmid that only contained the drug resistance protein. The last plate was treated with a plasmid containing the drug resistance protein and the CPAK2 protein. All three plates continue to be incubated.
January 28, 2008
We treated all the cells plates with the drug G-418. This drug will kill all the cells that do not present the drug resistance protein. This will tell us which cells present the CPAK2 protein.
January 25, 2008
So, the cells grew a little too well. There were several dead cells found when we took the plate out to split cells. On this day, the goal was to make 3 new plates with about 100,000 cells each. To get a rough estimate of how many cells were in the plate, we used a Hemacytometer to count cells. The cells were also treated with Trypan Blue. This is a dye that dead cells take in. We only wanted to count the live cells.
I also made medium. Our medium contains Pencillin-Streptomycin (to combat the growth of bacteria) and Newborn Calf Serum (contains growth factors that cause the cells to proliferate). These are both placed in DMEM.
Today was the first day that I have ever worked in a real lab. It was amazing. It was just a small step in the direction my major will take me. This semester I am going to be working with tissue cells. The goal is to change the DNA of the cells to make them produce a specific protein and a resistance to a drug. The drug resistance will allow us to kill all the cells that do not permanently produce the protein. Later, I will grow colonies of cells that produce the protein.
Today was a lesson in splitting the cells so that the cell line will continue. This has to be done every few day because the cells grow so fast and the medium will only support a specific number of cells before the medium becomes too acidic and the cells die. The next step is to take about 100,000 cells out of the millions in the plate and place them in one plate with medium. The next day a plasmid with the DNA needed for the protein and drug resistance will be introduced to the cell plate. It is expected that only about 100 of the 100,000 cells will permanently take the DNA into their own. The others will just discard the DNA as foreign.
The wetland flowers have been hit by the frost, and they look pretty drab now. Currently work is consisting of writing up the report and I am looking forward to working in the wetland next semester.
We left Freed at about 6 that morning to got to the Bat Conference near Nashville. Our group did well and we learned a lot form the 10 different presentations that were given.
Me and Tristan went out to the site. There was a little water near but we decided to go in anyway. We were only able to get to 7 trees and there wasn’t any bats in any of the trees.
