Nematode Growth Medium Culture Media

Molecular Biology

Store powdered media at RT. Opened bottles should be capped tightly and kept in a dark, low humidity environment. Prepared media should be kept at 4°C and used within a short period of time.

Light tan, homogeneous, free flowing powder

Light tan, incomplete

Light tan, clear, complete

7.0

Dissolve 23.005 grams in 973ml of ddH2O. Heat with stirring until the agar dissolves (cholesterol will typically remain undissolved). Dispense into appropriate containers, loosen caps and autoclave for 15 minutes at 121ºC (15psi).

Once the N1000 solution has cooled to 58°C, place flask(s) onto stir plate(s). Maintain temperature at 58°C. Add the following
a) Sterile 1M Phosphate buffer*, pH 6.0: 25ml/liter N1000 solution
b)Sterile 1M CaCl2: 1ml/liter N1000 solution
c) Sterile 1M MgSO4: 1ml/liter N1000 solution Allow the solution to mix for five minutes before starting to pour plates to ensure that a homogeneous mixture has been achieved. *1M Phosphate buffer is prepared by dissolving 108.3g KH2PO4 and 35.6g K2HPO4 in ddH2O. QS to 1L).

Components shown as g/liter:
Agar 17.5
Sodium Chloride 3.0
Peptone 2.5
Cholesterol 0.005
Total: 23.005g/L

Culture Media; Media-Nematode