Guidance for Preparing Standard Operating Procedures (SOPs)

Selanastrum B23.0
Rev. #: 1
Date: 12/99
Page 5 of 8
*
weigh out 100.0 mg and dilute to 100 ml. Add 10.0 ml of this solution to
Stock #3.
**
weigh out 220.0 mg and dilute to 100 ml. Add 10.0 ml of this solution to
Stock #3.
***
weigh out 60 mg and dilute to 100ml. Add 10.0 ml of this solution to
Stock #3.
b. Prepare macronutrient stock solutions by adding the chemical to 1L Super-Q water
in a volumetric flask.
1. Stock Solution #4: 36.76 g CaCl 2 .2H 2 O
2. Stock Solution #5: 36.97 g MgSO 4 .7H 2 O
3. Stock Solution #6: 12.60 g NaHCO 3
4. Stock Solution #7: 8.71 g K 2 HPO 4
5. Stock Solution #8: 85.01 g NaNO 3
c. Add 1 ml of each stock solution per liter of medium, except add 2 ml of stock
solution #2. Use a sterile pipette for each stock solution and dispose of after use.
d. Keep all stock solutions, except #3, in the dark at room temperature. Keep stock
solution #3 at 4 o C. Retain stock solutions for no longer than 24 months.
5.3 Starting an Algal Culture
a. Use a 2 L Erlenmeyer flask to prepare a 1.5 L batch of MBL medium. Add 1500 ml
of Super-Q water to the cool oven-dried flask. Add 1.5 ml of each stock except #2,
using a sterile 2.0 ml pipette for each solution. Add 3 ml of stock solution #2.
b. Inoculate the MBL medium with a slightly visible amount of inoculum from an agar
slant using a sterile inoculating loop and aseptic technique. An alternative
inoculum can be used by transferring 1-2 ml of a previous culture maintained at 4 o C
to the newly prepared mMBL medium.
c. Place a sterile pipette in a flask with a beveled syringe needle fixed to the pipette
tip. Attach to a filtered air source. This is done to prevent the algae from settling
and to provide adequate gas exchange. Stopper the culturing flask with a
gauze/cotton plug.
d. Cultures are kept at 25 + 2 o C (or at ambient temperature) at a light intensity of
approximately 400 ft-c. Lighting is continuous, i.e., 24 hours per day.
QA/G-6 D-5
March 2001