BBM & EBBM: for isolating photobionts and algae from BSC; this is a revised media protocol from other BBM protocols
To make the stock solutions:
Soln. # Chemical Weight Vol. ddH2O
Solution 1: K2HPO4 1.875 g 250 mL
Solution 2: KH2PO4 4.375 g 250 mL
Solution 3: MgSO4-7H2O 1.875 g 250 mL
Solution 4: NaNO3 12.5 g 500 mL
Solution 5 CaCl2-2H2O 0.625 g 250 mL
Solution 6: NaCl 0.625 g 250 mL
Solution 7: EDTA-Na4 5.0 g 100 mL
Solution 8: KOH 3.1 g 100 mL
Solution 9: FeSO4-7H2O 0.498 g 100 mL
H2SO4 (1.84 g/ 1 mL) add 100 uL
Solution 10: ZnSO4-7H2O 0.353g 25 mL
Solution 11: MnCL2-4H2O 0.058g 25 mL
Solution 12: CuSO4-5H2O 0.063 g 25 mL
Solution 13: Co(NO3)2-6H2O 0.20 g 25 mL
Solution 14: Na2MoO4-2H2O 0.048 g 25 mL
Use recipes for vitamin mix and trace elements that are explained above for our lab
*For longevity of the stock solutions it is advised that you autoclave them, however you can use them without autoclaving but they will only last about 3 months before expiring*
BBM: to make 1 L of media:
- Fill a 2 L Erlenmeyer flask with 940 mL of ddH2O
- Add 10 mL of solutions 1-6 to the flask
- Add 1 mL (1000 uL) of solutions 7, 8, 9 to the flask
- Add 100 uL of solutions 10-14 to the flask
- If making solid media add 15g of agar to the media
- Autoclave on P1 (less time liquid autoclave setting)
EBBM:
Do the same steps as above and:
- Add 30 mL of solution #4 instead of 10 mL
- Add 1 mL of Trace elements to the flask
- Add 1 mL of Vitamin mix to the flask Cyanobacteria: fixes N2 from the air so to differentiate cyanobacteria from algae do this Leave out Solution #4 of EBBM recipe