NGM Agar

NaCl
peptone
agar
cholesterol (5mg/ml in EtOH)
H2O

3 g
2.5 g
17 g
1 ml
975 ml

Autoclave, cool to 55°C, then add the following (mix after each addition):

1M CaCl2
1M MgSO4
1M Potassium Phosphate pH 6.0		 1 ml
1 ml
25 ml

 

RNAi plates

To regular NGM medium add (per L):

50 mg Ampicillin
240 mg IPTG (1 mM)

 

Egg plates

(Adapted from Methods in Cell Biology 48, page 515)

Egg plates can be used to grow large amount of worms on plates, and are especially usefull to grow unc-119 mutants. The egg yolks support rapid bacterial growth.

  1. In a sterile bottle, mix the following:
  2. Heat the mixture to 60°C for 10 min (to inactivate the lysozyme).
  3. Add 50 ml of an O/N OP50 culture.
  4. Dispense onto 2% NGM-agar plates (~4 ml for 10 cm plates and 8 ml for 15 cm plates).
  5. Dry the plates, and keep at 4°C.

 

M9 Buffer

KH2PO4
Na2HPO4
NaCl
1M MgSO4
H2O

3 g
6 g
5 g
1 ml
1 L

 

S Medium

S Basal
1M Potassium Citrate pH 6.0
Trace Metals Solution
1M CaCl
1M MgSO4

 1 L
10 ml
10 ml
3 ml
3 ml

S Basal

0.1 M
0.05 M
1 ml
1 L		 NaCl
Potassium Phosphate pH 6.0
cholesterol (5mg/ml in EtOH)
H2O

Trace Metals Solution

5 mM
2.5 mM
1 mM
1 mM
0.1 mM
1L		 Disodium EDTA (1.86 g)
FeSO4·7H2O (0.69 g)
MnCl2·4H2O (0.20 g)
ZnSO4·7H2O (0.29 g)
CuSO4·5H2O (0.025 g)
H2O

Autoclave and keep in the dark.

 

Freezing Solution

NaCl
KH2PO4
Glycerol
1M NaOH
H2O

5.85 g
6.8 g
300 g
5.6 ml
to 1L

Autoclave, then add:

0.1 M MgSO4 3 ml

 

1M Potassium Phosphate pH 6.0

Make 1M solutions of K2HPO4 and KH2PO4.
Mix the two in the following amounts:
132 ml K2HPO4
868 ml KH2PO4