Media Prep

Summary

This video guides viewers through the process of preparing brain heart infusion agar in a laboratory setting. It covers the necessary steps, from mixing deionized water with brain heart infusion agar powder, to sterilizing the media in an autoclave. The process also involves stirring, cooling, and pouring the media into petri dishes, allowing it to solidify before storage. Key points include proper handling of lab equipment, ensuring correct temperatures, and the importance of sterile conditions to promote cell growth.

Takeaways

• 😀 Cells need specific environmental conditions (food, energy, temperature, humidity) to grow in a lab setting.
• 😀 Solid culture media is a mixture of agar and nutrients, used for growing cells in petri dishes.
• 😀 The preparation of brain heart infusion agar involves adding deionized water to a flask with agar powder.
• 😀 Always check the lot number and expiration date before using culture media.
• 😀 Use an analytical balance and tare the scale to measure the correct amount of brain heart infusion agar (5.2g).
• 😀 After adding the agar to the flask, mix with deionized water and stir the solution to break down clumps.
• 😀 Cover the flask with aluminum foil and use autoclave tape to seal it before autoclaving.
• 😀 Autoclaving sterilizes the media and helps dissolve the agar in the water. Run the autoclave at 121°C for 17 minutes.
• 😀 Once sterilized, allow the media to cool to about 45°C before pouring it into petri dishes.
• 😀 After cooling, the agar will solidify and turn opaque. Store the petri dishes upside down to prevent moisture condensation.

Q & A

What are the essential environmental conditions for growing cells in the lab?

Cells need specific environmental conditions such as proper food, energy, temperature, and humidity to grow effectively in the lab.

What is solid culture media, and what is it made of?

Solid culture media is a mixture of agar and nutrients, which is poured into petri dishes to create the ideal environment for cell growth.

What is the purpose of brain heart infusion agar in this procedure?

Brain heart infusion agar is used as a culture medium to grow cells, and its preparation is demonstrated in this exercise.

What should you check before using a batch of culture media?

Before using culture media, always check the lot number and expiration date to ensure it's still viable for use.

How much deionized water should be added to the graduated cylinder at the beginning of the preparation?

You should add 100 mL of deionized water to the graduated cylinder to start the media preparation.

What is the function of the stir bar in the 250 mL flask?

The stir bar is placed in the flask to help mix the media and dissolve the components as the solution is stirred.

How is the correct amount of brain heart infusion agar measured?

The brain heart infusion agar is measured by using an analytical balance, where 5.2g of the powder is placed into a weigh boat and then added to the flask.

What should be done if using a combination hot plate and stir plate during media preparation?

If using a combination hot plate and stir plate, make sure that the heat is turned off to avoid overheating the solution while stirring.

What is the purpose of autoclaving the culture media?

Autoclaving sterilizes the culture media by using heat and pressure, ensuring the removal of any contaminants and aiding in the dissolution of the agar in the water.

Why is it important to let the media cool to 45°C before pouring it into petri dishes?

Allowing the media to cool to around 45°C ensures it remains in liquid form, which can then be poured into petri dishes, but not too hot to avoid damage to the dish or contamination risks.