From Protists

Contents 1 Filtered 95% Seawater 2 Soil Extract Soln 3 Stock Soln 4 Medium

Filtered 95% Seawater

• Filter 950 mL of natural seawater.
• Add 50 mL of distilled water and mix.
• Autoclave for 15 minutes at 121C.

Soil Extract Soln

• Fill a 1-L beaker with soil to the 200 mL line.
• Add distilled water to the 600 mL line (400 mL of water).
• Cover with foil and autoclave for 2 hours.
• Allow it to cool and filter it through Whatman #1 filter paper.
• Distribute 100 mL of the extract into 4 different 250-mL media bottles.
• Label as "PE Soil Extract".
• Autoclave for 15 minutes at 121C.
• Store in 4C walk-in until needed.

Stock Soln

Mix the following ingredients:

NaNO3		20.0g
NaHPO4.7H2O		1.5g
Distilled H2O		100mL

Label as "PE Stock Soln" and store in 4C walk-in until needed.

Medium

Mix the following ingredients aseptically (under the hood) into the final media bottles:

Filtered 95% Seawater		950 mL
PE Soil Extract		50 mL
PE Stock Soln		1 mL

• Label the Media bottles "PE".
• Autoclave the mixture for 15 minutes at 121C.
• Store in Media cabinet.