Oct 05, 2022
Obtain LB agar plates (with antibiotics)
- 1XJTLU
Protocol Citation: An.Huang 2022. Obtain LB agar plates (with antibiotics). protocols.io https://dx.doi.org/10.17504/protocols.io.j8nlkw4b5l5r/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: October 05, 2022
Last Modified: October 05, 2022
Protocol Integer ID: 70849
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Abstract
LB plate is a fundamental laboratory consumable. This protocol helps make basic LB agar plates.
Weigh 40 g LB nutrient agar powder.
Note
If do not have LB nutrient agar powder, use this as an alternative:
25 g LB broth powder and 15 g agar.
Dissolve the powder in 1000 mL distilled water.
Note
Agar may not completely dissolve in water at room temperature. Use water bath to help dissolve. Aliquoting the solution into smaller flasks is recommended, because it makes the following autoclaving and plating procedure more convenient.
Autoclave the melting LB agar at 121 °C for 00:15:00 .
Safety information
Use the autoclave safely.
15m
When autoclaving procedure is finished, wait until the flask cools to around 40 °C , which will not hurt the palm.
Safety information
Be careful with hot steam.
(Optional) If the plate is required to contain a certain antibiotic, add a certain amount of antibiotics.
Here ampicillin is used as an example:
Add 1 mL 1000X ampicillin stock solution into 1000 mL melting LB agar. Mix well.
Note
Add antibiotics when melting agar has not solidified.
Note
Different antibiotics may have different stock concentration. The most common concentration is 1000X stock solution. Add antibiotics accordingly.
Pour melting agar into empty petri dishes until the melting agar has completely cover the bottom of the petri dishes. Avoid bubbles.
Note
15 mL of melting agar is recommended. Adjust the volume according to the petri dishes used.
Note
If bubbles emerged, use a sterile loop to pop it.
Leave the plates to cool and the lids ajar.
Wait until all agar plates solidified. Invert the plates and store at 4 °C 。.
Note
Store temperature should be adjusted according to the antibiotics added.