| Reagent | Amount (per 100 mL) |
|---|---|
| Schneider's Drosophila Medium | 22 mL |
| Lactalbumin hydrolysate | 450 mg |
| Galactose | 130 mg |
| Sterile DI water | 68 mL |
| Gentamicin 50 mg/mL stock (optional) | 0.2 mL |
Filter sterilize (0.22 μm filter) and store at 4°C.
To prepare cBge medium, add 10 mL of heat-inactivated FBS and 1 mL of 100X pen/strep to 89 mL iBge.
<aside> ⚠️
iBge is very easily contaminated. Do not freeze cBge medium.
Bge cells are sensitive to the quality of FBS — several lots may have to be screened. Investigators using Bge cell lines have lost them due to unsuitable FBS.
</aside>
| Reagent | Amount (per 1 L) |
|---|---|
| NaCl | 2.8 g |
| Sodium gluconate | 1.5 g |
| KCl | 0.15 g |
| Na₂HPO₄ | 0.07 g |
| HCO₃ | 0.05 g |
| Glucose | 0.5 g |
| Trehalose | 0.5 g |
| Galactose | 0.5 g |
| Sterile DI water | 1 L |
| Reagent | Amount (per 1 L) |
|---|---|
| NaCl | 2.8 g |
| KCl | 0.15 g |
| Na₂HPO₄ | 0.07 g |
| MgSO₄ heptahydrate | 0.45 g |
| CaCl₂ dihydrate | 0.53 g |
| NaHCO₃ | 0.05 g |
| Sterile DI water | 1 L |
Filter sterilize (0.22 μm filter) and store at 4°C. To make CBSS+, add 1 g each of glucose and trehalose.
| Reagent | Amount |
|---|---|
| Trypsin | 50 mg |
| Hansen's buffered saline | 100 mL |
Filter sterilize (0.22 μm filter), freeze in 10 mL aliquots, and warm before using.
| Reagent | Amount (per 1 L) |
|---|---|
| CaCO₃ | 12.5 g |
| MgCO₃ | 1.25 g |
| NaCl | 1.25 g |
| KCl | 0.25 g |
| Sterile DI water | 1 L |
Pond water stocks are prepared in small Nalgene containers and stored on the supply rack in B7. Shake the stock bottle well before diluting, as the reagents and salts may have settled on the bottom.
Common working dilutions
| Pond water stock | Diluted into |
|---|---|
| 100 mL | 30 gallons dH₂O |
| 33 mL | 10 gallons dH₂O |
| 16.5 mL | 5 gallons dH₂O |
| 12 mL | 10 litres dH₂O |