NGM 1.2P Agar

Prepare a stock of phosphate buffer:

64.625g KH₂PO₄ (monobasic; FW136.09)

25.875g K₂HPO₄ (dibasic anhydrous, FW174.18)

to 500 ml with Milli-Q H₂O

Adjust pH to 6.0 with HCl or KOH and autoclave

This is called 1.2M KPO₄

To prepare 1L of NGM 1.2P Agar:

* Begin with approx. 700 mLs of Milli-Q H2O.

* Weigh out and add ingredients with constant stirring:

o NaCl 3g

o Bacto Peptone 2.5g

o Bacto Agar 16g

* Bring up to 1L with Milli-Q H₂O

* Autoclave

* Allow to cool in 50deg.C water bath

* Add:

• 1 M CaCl₂ (dihydrate; FW147.01) 1mL (final conc. 1mM)
• 1 M MgSO₄ (heptahydrate; FW 246.48) 1mL (final conc. 1mM)
• 1.2 M KPO₄ 25mL (final conc. 30mM)
• 5mg/mL cholesterol in 95-100% ethanol 1mL

Ian A. Hope (editor). C. elegans: A Practical Approach. Oxford University Press, New York, New York, 1999. Edited by S. Silverman, S. Skrovanek, 11/06.