Medium data

Search for medium no. = [1303].

Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

1303 UREAPLASMA MEDIUM

PPLO broth w/o Crystal Violet (BD-Difco 255420)	`2.1`	g
1% Phenol red solution (see below)	`200.0`	µl
Hipolypepton	`1.0`	g
Distilled water	`85.0`	ml

Mix components, adjust pH to 6.0-6.2. After autoclaving and then cooling to 50-55C, aseptically add the following solutions (filter-sterilized).

Bovine calf serum (heat-inactivated)	`5.0`	ml
1% Urea - 4% yeast extract solution (see below)	`10.0`	ml

1% Phenol red solution:
Dissolve 1.0 g phenol red first in 10-20 ml of 0.1 N NaOH. Then adjust volume to 100 ml. After filter-sterilizing, store at 4C.

1% Urea - 4% yeast extract solution:

Urea	`1.0`	g
Yeast extract	`4.0`	g
Distilled water	`100.0`	ml

Mix components, sterilize by filtration through a 0.22 µm filter and store at 4C.

Dispense the medium into sterilized plastic tubes (e.g., 5 ml medium in 15 ml of plastic tubes) and add 100-200 µl of culture solution. Incubate the cultures at 37C for 1-3 days. When the color of the medium turns from yellow to orange or red, subculture into the new medium. After color change, the strain can be stored at 4C for 1-2 weeks.