Clarified V8 agar

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Contents 1 Purpose 2 Ingredients 3 Instructions 4 Notes 5 References 6 Contributed by

Purpose

Sterol-containing medium that allows easy viewing of morphological features of oomycetes.

Ingredients

Ingredient
5x clarified V8 juice*	200 ml
dH2O	800 ml
Agar	15 g
Sucrose	1 g
Yeast extract	0.2 g

Instructions

1. Mix ingredients in 2 L flask.
2. Autoclave.

Notes

• To prepare clarified V8 juice*

1. Add 5 g CaCO3 to 340 ml V8 juice in a beaker containing a stir bar.
2. Stir for at least 15 m.
3. Aliquot into 50 ml Falcon tubes, making sure the tubes will balance in centrifuge—either by weighing or adjusting volume to graduations on tube.
4. Spin tubes in the benchtop centrifuge at 4000 rpm for 15 m.
5. Transfer clarified V8 not needed to 50 ml aliquots for storage at -20°C.
6. Label containers “5X V8” and note volume on container (e.g. 50 ml).
7. Place in -20°C.

References

• Tuite, J. 1969. Plant Pathological Methods: Fungi and Bacteria. Burgess Publishing Company, Minneapolis, MN. 239 pp.

Contributed by

From the Virginia Tech Mediabook; Orignially created by Robert Wick; contributed by Mary Ann Hansen