Standard Operating Procedure: 10L Nannochloropsis Subculture

To prepare a 10L total culture of Nannochloropsis, mix 8L of prepared media (Artificial Seawater + Nutrients) with 2L of active microalgae culture (Optical Density, OD > 1.0).

1. Inventory & Material Requirements

   • Microalgae Starter: 2L of Nannochloropsis culture.

   • Artificial Seawater (ASW) Mix: 500g total pack (Yields up to 14.5L of media / Cost: RM10.00).

   • Component A (Granular): 475g

   • Component B (Granular): 25g

   • Nutrient Mix: 100ml total pack (Yields up to 50.0L of media / Cost: RM50.00).

   • Base-A (Foliar): 50ml

   • Base-B (Foliar): 50ml

2. Formulation Per 1 Litre of Finished Culture

   To prepare each single litre of the final Nannochloropsis culture, combine the ingredients in the following sequence:

   • Water Base: Measure 1L of distilled water.

   • Salinity (ASW-A): Add 33.0g of ASW Component A and mix until completely dissolved.

   • Salinity (ASW-B): Add 1.7g of ASW Component B to the same mixture and stir until fully dissolved.

   • Nutrition (Base-A): Add 1ml of Nutrient Base-A and mix well.

   • Nutrition (Base-B): Add 1ml of Nutrient Base-B and mix well.

   • Inoculation: Inoculate the media by adding 200ml of active microalgae culture.

3. Batch Scaling for 10L Total Target Volume

   To scale up to your 10L final volume (requiring 8L of fresh media and 2L of inoculum), use these total quantities:

   • Distilled Water: 8.0 Litres

   • ASW Component A: 264.0 grams

   • ASW Component B: 13.6 grams

   • Nutrient Base-A: 10.0 ml (range from 1 to 4ml/L)

   • Nutrient Base-B: 10.0 ml (range from 1 to 4ml/L)

   • Microalgae Inoculum (OD > 1.0): 2.0 Litres

NOTE: Please refer to text instructions above. The image instructions is only for reference.