Medium data

Search for medium no. = [924].

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

924 PEAT MEDIUM 2 FOR METHANOBACTERIA

Major metals (see Medium No. 923)	`10.0`	ml
Trace metal 2 solution (see below)	`1.0`	ml
Distilled water	`1.0`	L

Mix components thoroughly, bring to a boil for 5 min, then, cool down under a stream of a N₂-CO₂ (80:20, v/v) gas mixture. Distribute the medium into Balch-type culture tubes (5 ml each) under the same gas mixture, seal with thick blue butyl rubber stoppers and autoclave.
To complete the medium, add the following solutions to each of culture tubes (containing 5 ml medium) from sterile anaerobic stocks (autoclaved or *filter-sterilized):

83 mM TiNTA solution* (see Medium No. 923)	`0.05`	ml
1.0 M MES solution* (pH 7.45)	`0.10`	ml
Vitamin solution* (see Medium No. 923)	`0.05`	ml
1% Yeast extract solution	`0.01`	ml
50 mM Coenzyme M solution	`0.025`	ml
10 mM Sodium acetate	`0.015`	ml
4 mM Na₂S·9H₂O solution*	`0.05`	ml

Pressurize the tubes to 70 kPa H₂-CO₂ (80:20, v/v) and allow to stand for at least 7 hr. Check the final pH of the medium to be about 5.7.
Trace metal 2 solution:

Trace metal solution 1 (see Medium No. 923)	`1.0`	L
EDTA·2Na	`37.23`	g

Adjust pH to 7.0 with NaOH.