Continued Work

              Throughout the weeks we’ve had in the lab, we have worked on a variety of things. We have been thoroughly sterilizing the lab, making brand new bacteria plates with kanamycin for future experiments, taking inventory, cleaning, and making broth. We did an experiment dealing with liquid culture and later recorded our results in a CFU assay. We then analyzed the results and tried to analyze if we did the experiment right or not. The liquid culture we grew turned out good, but we believe our experiment failed due to possible contamination or maybe having too much kanamycin on our plates. We are going to conduct another experiment soon to see if we did, in fact, contaminate or alter our experiment in any way. The purpose of our experiment was to figure out the optimal time to induce our bacteria with the turkey IL8 gene. By doing the CFU assay, we wanted to see when we have the optimal number of bacteria in our culture so we can induce them to produce the protein. We had to use the spectrophotometer every 30 minutes to see how much bacteria were in the liquid culture. We started the experiment at 7:30 in the morning and we had to examine the number of bacteria every 30 minutes until 3:00 p.m. We had to take the bacteria out of our samples and streak them onto plates, resulting in more than 100 plates. By doing this the first time, we possibly contaminated the plates since there were so many. I feel as if I have caught up a tremendous amount since we first started working in the lab, and I am very excited to see if our experiment will finally work the second time. 

Work in the Lab

We’ve learned a lot since we first got in school. Working in the Biotechnology lab is something eye opening and it makes you feel more professional working and knowing what you have to do. We learned how to properly sterilize our work environment before conducting an experiment or handling bacteria to avoid contamination. We’ve been working with E. coli bacteria recently (E. coli HB101 to be exact). We first learned how to properly handle bacteria and make sure not to breathe in the Agar powder and to handle the plates carefully to avoid exposing the bacteria. Agar is used as bacteria food in order to help them grow and make the colonies we need. Our first experiment was trying to make the bacteria glow when exposed to a UV light. By doing so, we needed to insert a gene into the bacteria found in jellyfish. We wanted to see if we could add the gene into the bacteria through heat shocking. We did not know for sure if it would work so we did the experiment. We had to keep the bacteria very cold to keep them sealed, and then we’d expose them to heat to make them swell up and expose pores, which would allow us to insert the GFP gene that will allow them to glow. After doing that, we had to put the bacteria back into the cold environment to make them seal up and trap the gene inside. The plates we had streaked had arabinose to help the bacteria activate the GFP gene and hopefully illuminate the colony. All of this had to be done in about 10 or 15 seconds, so there was no room for error. When we got back the next day, we had glowing colonies.

Then we took those bacteria colonies and streaked them on 4 different plates to see which one had growth and if they could glow. Our plates were -pGLO LB, -pGLO LB/amp, +pGLO LB/amp/ara, and +pGLO LB/amp. We wanted to see which plate grew the bacteria and most importantly, made them glow. We found out that the bacteria in the +pGLO LB/amp/ara plate grew and could glow, so we took the bacteria from that and streaked them onto a separate plate to see if they could still grow and we found out that our experiment worked.

First Post

     Although I did not personally go to NC State on the trip with my fellow classmates, I did go out of town. I went to Nags Head with my girlfriend and her family for the week and we stayed at a condo in Manteo. We went to the beach and saw lots of "sea butterflies" which are little groups of slugs that are harmless but can be a nuisance. After that, we went back to the condo and played pool in a clubhouse that comes with the condo.  After that, we went walking through downtown Manteo while playing Pokemon Go, and we ended up walking more than 3 miles in about 2 hours. I did want to go on the trip to Raleigh but this vacation was planned already and missing out may complicate starting right off the bat but in the long run I can look forward to my classmates helping me and working with me to be able to be proficient in this course.