Making Media and LB broth
June 5th, 2018
Today’s excitement did not stop at plating the kefir… our group got to make our very own LB broth and
media to use for our future tests.
media to use for our future tests.
For those who don’t know, Luria Bertani (LB) Broth is a liquid medium that helps bacterial growth.
We mix our kefir, plain yogurt, and flavored yogurt in it so that the bacteria they contain can grow
overnight. Media is somewhat similar to LB; the solution used to make media is placed in an autoclave
(a high pressure, high temperature cooker) for an hour so that the media and containers become sterile.
Unlike LB, media will cool down and solidify.
We mix our kefir, plain yogurt, and flavored yogurt in it so that the bacteria they contain can grow
overnight. Media is somewhat similar to LB; the solution used to make media is placed in an autoclave
(a high pressure, high temperature cooker) for an hour so that the media and containers become sterile.
Unlike LB, media will cool down and solidify.
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.
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.
Theresa stirring the 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.
Theresa stirring the LB Broth:
Comments
Post a Comment