# J988: MPOB MEDIUM


## Main sol. J988

0.53	g	Na2HPO4	
0.41	g	KH2PO4	
0.3	g	NH4Cl	
0.11	g	CaCl2 x 2 H2O	
0.1	g	MgCl2 x 6 H2O	
0.3	g	NaCl	
1	ml	**Trace element solution SL-10**	
1	ml	**Selenite-tungstate solution**	
0.4	g	Yeast extract (BD-Difco)	
0.5	mg	Resazurin	
4	g	NaHCO3	
10	ml	**Trace vitamins**	
2.3	g	Glycerol	
0.5	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Mix ingredients except Trace vitamins, NaHCO3 and Na2S x 9H2O. Separately autoclave NaHCO3 and Na2S x 9H2O (4 and 5% solutions, respectively) under a N2-CO2 atmosphere. Anaerobically and aseptically add the NaHCO3 and Na2S x 9H2O solutions, and filter-sterilized Trace vitamins to the medium. Check final pH to be 7.0 - 7.2. Pressurize the inoculated bottles to 70 kPa N2-CO2 (4:1, v/v).


## Trace element solution SL-10

10	ml	HCl (25%, 7.7 M)	
1.5	g	FeCl2 x 4 H2O	
70	mg	ZnCl2	
100	mg	MnCl2 x 4 H2O	
6	mg	H3BO3	
190	mg	CoCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
24	mg	NiCl2 x 6 H2O	
36	mg	Na2MoO4 x 2 H2O	
990	ml	Distilled water	

1. Dissolve FeCl2 x 4H2O first in HCl. Then dilute in distilled water and add the remaining salts.


## Selenite-tungstate solution

0.5	g	NaOH	
3	mg	Na2SeO3 x 5 H2O	
4	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled water	


## Trace vitamins

2	mg	Biotin	
2	mg	Folic acid	
10	mg	Pyridoxine hydrochloride	
5	mg	Thiamine HCl	
5	mg	Riboflavin	
5	mg	Nicotinic acid	
5	mg	Calcium pantothenate	
0.1	mg	Vitamin B12	
5	mg	p-Aminobenzoic acid	
5	mg	Lipoic acid	
1000	ml	Distilled water