BOLD’S BASAL MEDIUM (MODIFIED)

BOLD’S BASAL MEDIUM (MODIFIED)

This medium is highly enriched and is used for many of the green algae. Reference: Stein, J. (ED.) Handbook of Phycological methods. Culture methods and growth measurements. Cambridge University Press. 448 pp.

NO	STOCK	STOCK SOLUTION	ml/Litre
1	KH2PO4	8.75 g/500 ml	10 ml
2	CaCl2.2H2O	1.25 g/500 ml	10 ml
3	MgSO4.7H2O	3.75 g/500 ml	10 ml
4	NaNO3	12.5 g/500 ml	10 ml
5	K2HPO4	3.75 g/500 m	10 ml
6	NaCl	1.25 g/500 ml	10 ml
7	Na2EDTA. 2H2O	10 g/L	1 ml
8	KOH	6.2 g/L
9	FeSO4.7H2O	4.98 g/L	1 ml
10	H2SO4 (concentrated)	1 ml/L
11	Trace Metal Solution	See below	1 ml
12	H3BO3	5.75 g/500 ml	0.7 ml

Adjust the pH to 6.8. OPTIONS: For 10% BBM, use 100 ml of 100% BBM/Litre of distilled water. The addition of 5 ml of soil extract is also beneficial to some algae.

Trace Metal Solution:

No	Substance	g/Litre
1	H3BO3	2.86 g
2	MnCl2.4H2O	1.81 g
3	ZnSO4.7H2O	0.222 g
4	Na2MoO4.2H2O	0.390 g
5	CuSO4.5H2O	0.079 g
6	Co(NO3)2.6H2O	0.0494 g

Dissolve each of the above substances separately prior to adding the next on the list.