MBL Medium - IBEMicrobeHub/culturing GitHub Wiki

For dinoflagellates.

Materials:

  • dH2O

Table 1

MBL Medium g in 1L of stock mL in 1L of MBL medium
1. CaCl2·2H2O 36,76 g 1 mL
2. MgSO4·7H2O 36,97 g 1 mL
3. NaHCO3 12,60 g 1 mL
4. K2HPO4 8,71 g 1 mL
5. NaNO3 85,01 g 1 mL
6. Na2SiO3·9H2O 28,42 g 1 mL
7. Na2EDTA 4,36 g 1 mL
8. FeCl3·6H2O 3,15 g 1 mL
9. Metal Mix See recipe below (1) 1 mL
10. Vitamin Stock See recipe below (2) 3 mL (1 mL / vitamin)
11. Tris Stock 250,00 g 1 mL

(1) Metal mix:

Table 2

9. Metal Mix g in 1L of stock
CuSO4·5H2O 0,010 g
ZnSO4·7H2O 0,022 g
CoCl2·6H2O 0,010 g
MnCl2·4H2O 0,180 g
Na2MoO4·2H2O 0,006 g

(2) Vitamin stock:

Table 3

10. Vitamin Stock g in 1L of stock mL in 1L of MBL medium
Cyanocobalamin (B12) 0,0005 g 1 mL
Thyamine HCl (B1) 0,1000 g 1 mL
Biotin 0,0005 g 1 mL

Method:

  1. Prepare all the stock solutions with dH20, and store them in the refrigerator.
  • Stocks 1-8 & 11: Add the quantity of each component (table 1) and fill each one up to 1L of dH2O.
  • Metal Mix: Add each constituent separately to 750 mL of dH2O (table 2), fully dissolving between additions. Finally, make up to 1L with dH2O.
  • Vitamin stock: Add the quantity of each component (table 3) and fill each one up to 1L of dH2O.
  1. Prepare the final MBL medium:
  • Add 1 mL of each stock solution (1-11) to 1L dH2O.

For species that cannot use nitrate substitute 1 mL of NH4Cl made up to 5,4 g/L H2O.

  • Adjust the pH to 7,2 with HCl.
  • Autoclave at 121ºC (15PSI for 15 min).