Medium data

Search for medium no. = [1355].

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

1355 MODIFIED M9 MEDIUM

Na₂HPO₄	`4.33`	g
NaH₂PO₄	`2.69`	g
NaCl	`20.0`	g
KCl	`0.13`	g
NH₄Cl	`0.31`	g
Distilled water	`835.0`	ml

Mix components thoroughly and autoclave under a N₂ atmosphere. After cooling, aseptically and anaerobically add the following solutions (autoclaved or *filter-sterilized and stored under a N₂ atmosphere):

Modified trace vitamins solution* (see below)	`10.0`	ml
Modified trace minerals (see below)	`5.0`	ml
10% Na₂CO₃ solution	`40.0`	ml
1 M Sodium acetate solution	`10.0`	ml
Fe NTA solution (see below)	`100.0`	ml

Modified trace vitamins solution:

Trace vitamins (see Medium No. 197)	`1.0`	L
Sodium ascorbate	`5.0`	mg
myo-Inositol	`5.0`	mg
Choline chloride	`5.0`	mg
Nicotinamide	`5.0`	mg

Modified trace minerals:

Nitrilotriacetic acid	`1.5`	g
MgCl₂·6H₂O	`3.0`	g
MnCl₂·2H₂O	`0.5`	g
NaCl	`1.0`	g
FeCl₂·4H₂O	`0.1`	g
CoCl₂·6H₂O	`0.1`	g
CaCl₂·2H₂O	`0.1`	g
ZnCl₂	`0.1`	g
CuCl₂·2H₂O	`0.01`	g
AlCl₃	`0.01`	g
H₃BO₃	`0.01`	g
Na₂MoO₄·2H₂O	`0.01`	g
Distilled water	`1.0`	L

Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH. Then proceed to add minerals. Adjust final pH to 7.0.
Fe NTA solution:

NaHCO₃	`1.64`	g
Nitrilotriacetic acid trisodium salt monohydrate	`2.75`	g
FeCl₃·6H₂O	`2.7`	g

Firstly, dissolve NaHCO₃ in 80 ml distilled water. Then, add nitrilotriacetic acid trisodium salt and FeCl₃·6H₂O to the solution, and bring volume to 100 ml with distilled water. Adjust pH to 7.0 with NaOH. Centrifuge the solution and filter-sterilize the supernatant. Store the solution under a N₂ atmosphere.