First thing to do when the SH media arrived was to reconstitute 1L of it, and get my samples growing. However, sterilization was going to be a big issue for us, as we had no autoclave. Fortunately, we were able to make do with a makeshift autoclave - Dr. B's pressure cooker.
Below is the methods write-up:
Wednesday, 10/30
Using a tin weigh boat and metal spatula, the entire contents of the SH Media bottle was massed on an electronic balance and determined to be 32.0 grams. If 32.0 g can be reconstituted to from 10L, then it follows that to make 1L would 3.2 g. Accordingly, all powdered media was placed back into the original container except for 3.2 g, and stored at 2-8˚C. To a 2 L Erlenmeyer flask was added 3.2 g of SH Media and 1 L of distilled water, and swirled until dissolved. 150 mL of the solution was apportioned into four 250 ml Erlenmeyer flasks each; the large Erlenmeyer flask containing the remaining solution was covered over the opening with tinfoil and left to stand at room temperature.
Then, the 250 ml flasks containing the media were each covered with tinfoil at their openings and placed into a small-sized safety pressure cooker. The pressure cooker was then sealed and placed onto a hot plate, with the heat setting turned on high. The pressure cooker was heated until whistling; the heat was then turned down to 8 and let sit for 20 minutes. After 20 minutes, the heat was turned off, and the flasks were left to cool.
Friday, 11/1
A jar of samples were taken out of the greenhouse. Forceps were used to pick up several colonies by transferring them to a petri dish, and then isolating four separate fronds (each their own colony). One colony was transferred to their respective flask, and labeled. The flasks were then taken to the greenhouse, covered lightly with tinfoil, and exposed to normal intervals of sunlight and darkness.
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