To isolate micro-organisms for the HeadsUp project, LB agar plates were made according to prot-001. There was a minor change because instead of cooking the agar in the pressure cooker for 30 minutes the solution was boiled under atmospheric pressure for an hour. The 500 ml of LB agar was poured into 12 Petri dishes. 3 of these plates were used in this experiment. The first plate was put with the lid open in a room for 30 minutes. On the second plate, a thumb was pushed in the center. The third plate functioned as a control and was never opened. The results are shown in figure 1.

Figure 1: pictures of the LB agar plates. From left to right, the control, 30 minutes opened, and a thumb pressed on the agar.

The results show that the control was full of growing bacteria. These bacteria were found on the surface and inside the agar. This is an indication that sterilization did not yield the wanted result. From my studies, I know that we always autoclave at 121 degrees of Celsius. The control suggests that 100 degrees are not enough. This "background" of bacteria as seen in the control was also present in the other two plates. These background bacteria are so numerous that it becomes difficult to isolate any bacteria from the other two plates. Moreover, the thumb has two many bacteria that are close together. This makes it also unfit to isolate micro-organisms. The plate that was left open for 30 minutes has a nice variety of bacteria and fungi. It seems that using this technique to find different species of micro-organisms is the way to go.

The next challenge was to get the temperature of the agar to 121 degrees or more. Using a pressure cooker the boiling temperature can be increased. Therefore, a pressure cooker was acquired (see figure 2). The pressure cooker reaches a pressure of 200.000 Pa, which means the boiling point reaches around 121 degrees.

Figure 2: A picture of the acquired pressure cooker.

The pressure cooker was immediately put to the test! 500ml of LB agar was made (see prot-001) and poured into 12 Petri dishes. 4 plates were used in the next experiment. The first plate was used as a control and was unopened. The other 3 plates were opened for 30 minutes in three different rooms (the bedrooms of my roommates). The results are shown in figure 3.

Figure 3: pictures of the second batch of LB agar plates. The top left depicts the control, the other plates are left open for 30 minutes inside my roommate's bedroom.

The results indicate that Tim has the cleanest room and that DJ has the dirtiest. The control shows no growth suggesting that the pressure cooker worked perfectly as an autoclave. The next step is to collect and isolate strains that are useful for project heads-up. Figure 4 shows three of the resulting isolates.

Figure 4: The first 3 isolates from project HeadsUp.

The results show that the isolation was a succes. The left plate does have a minor contamination.