June 15, 2017

One of our first meetings as University of Delaware’s iGem team members this summer was to go over safety in the lab. Sam one of our team’s leaders thoroughly explained micro pipetting and using aseptic technique to transfer bacterial cultures from agar growth plates, a process we will later use in our experiments. Aseptic technique, as Sam explained, is a sanitary way of moving bacterial so that their environments don’t become contaminated by foreign substances. Even clean air carries microbes!

Figure 1

Figure 1 shows eager team members listening in the lab (space provided by the University of Delaware) to Sam’s lecture on aseptic technique. This is where much of our experimenting will occur for our project.

Figure 2

In Figure 2 a team member demonstrates the proper way to handle a pipette with a sterile tip, holding it vertically and out of harm’s way (gloves and safety goggles normally worn in the lab were not necessary for this demonstration as no harmful chemicals were used, just water for demonstration purposes).

Figure 3

Figure 3 is the work space of a group member showing the proper way to clean up one’s space after working in the lab. Notice that it was wiped down with Bacdown, an antibacterial disinfectant and all beakers are tightly closed. In addition, the Bunsen burner is detached from the gas nozzle and the space is clear of debris.

Figure 4

Figure 4 shows Sam, one of our team leaders, explaining to fellow team members the importance of keeping the box that houses sterial pipetting tips. He explained that the least amount of exposure to air is essential to ensure that mold or other contaminates do not interfere with experimenting.

June 29, 2017

On June 29, 2017 our team met to go over proper streaking technique. This means that we learned how to properly transfer a bacteria culture from one petri dish to another using sterile loops. The point of transferring cultures from one petri dish to another is in order to grow new cultures from one similar culture to obtain a greater volume for experimenting on. In addition, we learned that we do not want too many clusters of bacteria growing on top of each other because it would make it hard to identify individual cultures. me. It's easy.

Figure 1

In Figure 1 a team member demonstrates the proper way to handle a sterial loop when transferring bacteria. It is essential that nothing touch the loop in order to ovoid contamination of the petri dish.

Figure 2

Figure 2 is a black board drawing that a student made to help other team members understand the proper way of streaking. Quadrant 1 (top left of petri dish) will have the most condensed amount of bacteria while quadrant 4 will have the most diluted and spaced out colonies.

Figure 3

These are proper labeled petri dishes before they have been streaked with bacterial cultures.

Figu

This is a petri dish after some days of incubation that shows bacterial growth. Notice how the colonies grow further apart on the left compared to the heavily streaked right most quadrant.

Figure 5

Here is the team today all working hard and wearing goggles, gloves and long pants, the proper safety attire.

Attendance:

Yianni
Cassandra
Maddie
Adam
Emma
Ian

June ,5 2017

Meeting Agenda:

Sterile technique, taught by Tisa, and first day inside the lab

Attendance (rough)

Seth Martinez
Kira Dzedzy
Ioannis (Yianni) Zerefos
Kathleen Wright
Madlen Can
Carissa Walkosak
Dan Owens
Kelsey Murray

June 6, 2017

Meeting Agenda:

Plasmid Construct Discussion

Attendance (rough)

Kathleen Wright
Sara Leung
Maddie Callahan
Madlen Can
Kira Dzedzy
Ioannis (Yianni) Zerefos

June 7, 2017

July 9, 2017

Today’s meeting focused on resuspending the DNA sequences obtained from iGem, and measuring the concentrations to verify them. Following iGem recommendation we resuspended the DNA in 10 𝛍l of DI water, before adding 50 additional 𝛍ls to the solution. We used a spectrophotometer to analyze the concentration of this 60 𝛍l solution, with the following results:

Emma Oskar and Sam one on one meeting.

June 9, 2017

Sterile technique, Second session

Attendance

Seth Martinez
Sara Leung
Madlen Can
Kira Dzedzy
Ioannis (Yianni) Zerefos
Maddie Callahan

June 23, 2017

Plate Streaking Practice

Attendance:

Kira
Yiauni
Madlen
Sarah

The results of this preliminary test showed a concentration below what was expected, to be sure and eliminate any error, further trials were run in an experiment. In the first section, three wells were resuspended and each moved into three centrifuge tubes before being diluted to a total volume of 60 𝛍ls. In the

second section, six total wells were resuspended and

moved into three centrifuge tubes at two wells apiece, each tube was diluted to a total volume of 120 𝛍ls. And in the third section a similar procedure was carried out, simply resuspending and adding three wells worth of dna to to each tube before diluting each tube to a total volume of 180 𝛍ls. A diagram of the wells taken and the corresponding results of the experiment can be seen below (Note: The second of the 120 𝛍l tubes should be lower in concentration, accounting for a minor error):

(here post the photos of the design from the board and of the long results printout sheet)

July 10, 2017

Figure 1

This is a package of dehydrated DNA cells which is what we resuspended in DI water.

Figure 2

Two team members: Yanni and Sarah work together.

Figure 3

Yanni writes on the board while Sam dictates our data so that the whole team is coordinated.

Figure 4

This is a diagram Sam made so that team members knew which wells they were extracting DNA from and how much they were res-suspending in water.

Figure 5

On the board is written the volumes and coordinates for the wells that the DNA was extracted from so that when the tests went through the Spectrophotometer we knew which data was for which test.

Figure 6

Maddie is bummed that when she was extracting DNA she pressed too hard on the bottom of the well with a pipette so that the tip bent meaning that this test is invalid.

Figure 7

Ian and Sam use a Spectrophotometer to measure concentrations of resuspended DNA.

Figure 8

The Spectrophotometer that we use in lab.

Figure 9

These are the results from the Spectrophotometer for our DNA concentrations (see Figure 5 to tell which test # is for which concentration)

Today during our meeting we discussed and worked on logistics. We worked on the team t shirt, wiki and logistics for the Mid Atlantic meet up. We also looked at the results for the transformations that we ran yesterday.