LB Broth Agar Plates with Antibiotics

Bailey Clark

Published: 2022-06-28 DOI: 10.17504/protocols.io.kxygxz974v8j/v1

Abstract

The abstract will be added later.

Steps

Preparation For Autoclave

1.

Add 125mL of deionized water to a 250 mL Erlenmeyer Flask.

2.

Add 3.125g of LB Broth powder (25g per 1L of water) to the water in the Erlenmeyer Flask.

3.

Add 1.875g of agar to the water ( 1.5% of 125mL ) in the Erlenmeyer Flask.

4.

Stir on stirring plate for 0h 5m 0s .

5.

Cover the opening of the Erlenmeyer Flask with aluminum foil.

6.

Place a 10 mm strip of autoclave tape on the aluminum covering the opening of the flask.

Autoclaving

7.

Autoclave.

Equipment

ValueLabel
Electronic Table-Top AutoclaveNAME
EZ11PlusTYPE
TuttnauerBRAND
MAN205-0443001EN Rev SSKU
https://tuttnauer.com/laboratory-autoclaves/laboratory-research/benchtop-autoclaves/el-d-lineLINK

After Autoclaving

8.

After autoclaving, place the flask in the water bath at 60°C for 0h 15m 0s .

Equipment

ValueLabel
General Purpose Digital Water BathNAME
PolyScienceBRAND
WBE20A11BSKU
9.

Add 1µL of the antibiotic stock solution per mL of LB agar in the flask.

10.

Stir on the stirring plate for 0h 1m 0s .

11.

Pipette 5mL of the LB agar mix into 20 60 mm petri dishes.

12.

Leave the surface of the agar 1/4 uncovered in the laminar flow hood for 0h 30m 0s .

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