Zamanian Lab C. elegans recipes.
M9 buffer (4X stock)
| Reagent |
Amount for 4 L |
| KH₂PO₄ |
48 g |
| Na₂HPO₄ |
96 g |
| NaCl |
80 g |
| MQ H₂O |
up to 4 L |
- To make 4 L of 4X stock, measure all reagents into 3 L milliQ water in a 6 L flask and fully dissolve. Split the volume into two 2 L graduated cylinders and bring each to 2 L. Combine in the 6 L flask and aliquot into eight 500 mL aliquots.
- Autoclave on liquid cycle for 1 hour. Loosen the cap only a little, to maintain final volume.
- When the M9 is cool, add 2 mL of sterile 1 M MgSO₄ per 500 mL of M9 buffer.
Use autoclaved 2 L water bottles (1500 mL) to dilute to 1X for use in assays.
MgSO₄, 1 M (for M9 and S Medium)
- To make 500 mL, completely dissolve 123.24 g of MgSO₄ heptahydrate (FW 246.48) in 450 mL milliQ water from the carboy. Transfer to a graduated cylinder and bring up to final volume.
- Autoclave for 1 hour.
Potassium phosphate buffer, 1 M pH 6 (for S Basal/Medium and freezing solution)
| Reagent |
Amount |
| KH₂PO₄ |
136.1 g |
| KOH |
17.99 g |
| MQ H₂O |
up to 1 L |
Autoclave for 1 hour. Store in 500 mL aliquots.
S Basal
| Reagent |
Amount |
| NaCl, 5 M |
20 mL |
| Potassium phosphate buffer, 1 M pH 6 |
50 mL |
| Cholesterol, 5 mg/mL in ethanol |
1 mL |
| MQ H₂O |
930 mL |
Autoclave on liquid cycle for 1 hour. The solution will be cloudy.
K medium
| Reagent |
Amount |
| NaCl, 5 M |
5.1 mL |
| KCl, 1 M |
16 mL |
| CaCl₂, 1 M |
1.5 mL |
| MgSO₄, 1 M |
1.5 mL |
| MQ H₂O |
500 mL |
- Mix the pre-autoclaved salts above.
- Filter sterilize using a 0.22 μm filter.