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Document: LB medium | Last modified: June 04, 2005
LB Media (With Or Without Agar)

Final volume : 4.0 L

Bacto Tryptone : 40.0 g
(Difco, Cat. #0123-01-1)
Located in the chemical closet.

Bacto Yeast Extract : 20.0 g
(Difco, Cat. #0127-01-7)
Located in the chemical closet.

NaCl : 40.0 g
(Fisher Scientific, Cat #3624-05)
Located in the cupboard under the back bench.

If LB Agar is being made
Bacto Agar : 40 g (1%)
(Difco, Cat. #0140-01)
Located in the chemical closet.

Fill a 4 L beaker (obtained from the kitchen) with 2 L DDW. Put in a large stirbar.

Add ingredients for media without agar (in the order seen above). Let one ingredient dissolve before the next is added.

Adjust the volume to about 3.7 L. Bring pH up to 7.5 using 2 N NaOH (found by the pH meter).

Add DDW to the final volume (4 L) in a graduated cylinder (2 x 2L)

Aliquot into 500 ml bottles. Leave at least 1/5 of the bottle empty so the media won't boil out during the autoclaving.

If agar is needed, add the agar (example: if 10 volumes of 400 ml are made, each will recieve 4 g of agar).

Cover bottles with tin foil and a piece of autoclave tape with the room number.

Send to kitchen to be autoclaved.

Store on shelvess located by the door to the rear of the lab.

LB media with or without agar must be autoclaved right away otherwise it will quickly become contaminated. Do not make the media at the end of the day!
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