Mar 05, 2024
Making tetracycline LB agar plates
- Bonnie Evans1,2
- 1MRC Laboratory of Medical Sciences;
- 2Imperial College London
Protocol Citation: Bonnie Evans 2024. Making tetracycline LB agar plates. protocols.io https://dx.doi.org/10.17504/protocols.io.yxmvm388bl3p/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: March 05, 2024
Last Modified: March 05, 2024
Protocol Integer ID: 96160
Abstract
Making LB agar plates containing tetracycline for selecting strains carrying the pJC8 cosmid. Plates can be stored at 4 °C wrapped in aluminium foil for up to a month.
Making tetracycline LB agar plates
Making tetracycline LB agar plates
Autoclave or microwave LB agar from media room
Note
Contents of LB Agar:
37 g pre-mixed powder consisting of:
- 5 g peptone
- 10 g peptone from casein
- 10 g sodium chloride
- 10 g g agar-agar
1 L Sterile H2O
Make up 15 mg/ml tetracycline hydrochloride in sterile water
Retrieve tetracycline hydrochloride from -20 °C freezer
Weigh an empty Eppendorf
In the fume hood, add tetracycline hydrochloride to the Eppendorf
Weigh the Eppendorf, and calculate the difference to determine the weight (mg) of compound
To calculate sterile water (uL) to add = (compound mg / 15 mg/mL) x 1000 uL
Let LB agar cool to 55 °C
Dilute tetracycline hydrochloride 1/1000 dilution in LB agar (final conc: 15 ug/mL).
Using a serological pipette, dispense 35 mL agar into 90 mm Petri dishes. Leave to set.