Making Media Day II
Well, we were back in prep room making some more media. We used about 75 plates in one week which was surprising.
Meet our lab technician Richard! He was taught us how to make media and LB. Without him, we would not be able to carry out our experiments.
We followed the following processes to make media, LB, autoclave, and plate them.
Meet our lab technician Richard! He was taught us how to make media and LB. Without him, we would not be able to carry out our experiments.
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| Richard: The MVP |
We followed the following processes to make media, LB, autoclave, and plate them.
How to Make Media:
1. Measure 1,000 mL of distilled water in a graduated cylinder. Pour it into the glass pitcher.
2. Using the electronic scale, measure 25 g of the LB powder. Add it the dH2O.
3. Using the electronic scale again, measure 15 g of powder Agar. Add it to the pitcher.
4. Mix until the power is no longer visible. The liquid inside of the pitcher will resemble broth.
5. Tightly cover the pitcher with foil.
6. Place it in the autoclave for 1 hour. The solution will be heated at 121 degrees Celsius for 15 minutes.
The autoclave will heat and cool within the one hour cycle.
7. After the process is done, it is time to pour the media into the petri dishes.
8. Remove the foil from the pitcher and set up the petri dish on a paper towel. Some liquid media will spill.
9. Pour enough media in the bottom of each plate until the bottom is covered and it appears to be
darkish yellow in color. Keeping pouring the media into different petri dishes until there is no more left.
10. Let the plates solidify over night.
1. Measure 1,000 mL of distilled water in a graduated cylinder. Pour it into the glass pitcher.
2. Using the electronic scale, measure 25 g of the LB powder. Add it the dH2O.
3. Using the electronic scale again, measure 15 g of powder Agar. Add it to the pitcher.
4. Mix until the power is no longer visible. The liquid inside of the pitcher will resemble broth.
5. Tightly cover the pitcher with foil.
6. Place it in the autoclave for 1 hour. The solution will be heated at 121 degrees Celsius for 15 minutes.
The autoclave will heat and cool within the one hour cycle.
7. After the process is done, it is time to pour the media into the petri dishes.
8. Remove the foil from the pitcher and set up the petri dish on a paper towel. Some liquid media will spill.
9. Pour enough media in the bottom of each plate until the bottom is covered and it appears to be
darkish yellow in color. Keeping pouring the media into different petri dishes until there is no more left.
10. Let the plates solidify over night.
While the media was sterilizing in the autoclave, Nadine, Reika, and I made the LB broth.
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| Nadine pouring media |
How to Make LB Broth:
1. Measure 500 mL of distilled water in graduated cylinder. Pour it in a pitcher.
2. Using the electronic scale, measure 12 g of LB powder.
3. Mix the powder into the dH2O until it is dissolved.
4. The LB broth will be placed into a flask where the solution and container will be sterilized in the
autoclave before use.
1. Measure 500 mL of distilled water in graduated cylinder. Pour it in a pitcher.
2. Using the electronic scale, measure 12 g of LB powder.
3. Mix the powder into the dH2O until it is dissolved.
4. The LB broth will be placed into a flask where the solution and container will be sterilized in the
autoclave before use.
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| Reika Plating her Media |
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