Agar powder or LB broth powder
DI water
Petri dishes
Jar
Stir bar, stir plate
Autoclave, autoclave tape
Appropriate broth
Test tubes or petri dishes
Bunsen burner and striker
Inoculating loop
Parent culture
Incubator
1. Determine the volume of broth you would like to create and determine how much broth powder is required.
2. Place a piece of autoclave tape labelled with the broth type and date onto the jar you will be using.
3. Measure the appropriate amount of powder using a weigh paper and scoopula.
4. Place this powder in the jar. Place a stir bar in the same jar. Add the appropriate amount of DI water.
5. Mix the powder and water using the stir plate until all powder is dissolved.
6. Slightly unscrew the lid and run the jar on the liquid autoclave setting.
7. After autoclaving, place the jar in storage at room temperature.
1. Determine the volume of agar you would like to create and determine how much agar powder is required.
2. Place a piece of autoclave tape labelled with “Agar, date” onto the jar you will be using.
3. Measure the appropriate amount of powder using a weigh paper and scoopula.
4. Place this powder in the jar. Place a stir bar in the same jar. Add the appropriate amount of DI water.
5. Mix the powder and water using the stir plate until all powder is dissolved.
6. Slightly unscrew the lid and run the jar on the liquid autoclave setting.
7. After autoclaving, begin pouring plates. Add enough agar to entirely cover the bottom of the plate.
8. Leave the plates at room temperature overnight to allow them to set.
9. Store the plates with the lid down in at 4°C until use.
1. Gather materials. Into a test tube, transfer 10mL of LB broth using the automatic pipette. Label this test tube with the intended bacteria contents, your name, and the date of inoculation.
2. If you are using a liquid parent culture: transfer 200uL of the parent culture using a pipette into the new culture. Vortex sufficiently.
3. If you are using a solid parent culture: light the Bunsen burner. Flame the inoculation loop until it is red. From the parent culture, scrape off some growth. Place the loop inside the test tube and shake the loop to release cells.
4. Place test tubes in the shaker in the incubator at 200rpm. Grow overnight at 37°C.
5. After growth, store test tubes in an appropriate rack at 4°C.
1. Gather materials. Label the bottom of the petri dish (the side that contains the agar) with the intended bacteria contents, your name, and the date of inoculation.
2. If you are using a liquid parent culture: transfer 200uL of the parent culture using a pipette onto the new culture plate. Proceed to step 9.
3. If you are using a solid parent culture: light the Bunsen burner. Flame the inoculation loop until it is red. From the parent culture, scrape off some growth. Proceed to step 9.
4. Spread the fluid with the inoculation loop (liquid) or spread the inoculation loop (solid). Spread sufficiently around the full plate.
5. Plate the plates in the incubator at 37°C with the lid down overnight.
6. After growth, store petri dishes at 4°C.