Table 1. Transfection conditions for zebrafish PAC2 fibroblast cells
| Dish type | Cell number (×106) | Cell suspension (mL) | Total vol. (mL)a | DNA (μg) | Nanofectin diluentb (μL)c |
FuGENE HD (μL) | Opti-MEM/D-MEM (μL) |
|---|---|---|---|---|---|---|---|
| Four- or 24-well | ~2.5 | 0.17 | 0.5 | 0.75-1.5 | 50 | 2 | 50 |
| MatTek | ~1.5 | 1.0 | 3 | 3-6 | 100 | 8 | 200 |
| Petri dish, 35-mm | ~2.0 | 1.3 | 4 | 3-6 | 100 | 8 | 200 |
| Petri dish, 10-cm | ~4.0 | 3.3 | 10 | 20 | 250 | 20 | 500 |
| aTotal volume = 1:3 dilution of cell suspension obtained from dish of 100% confluent cells. | |||||||
| b150 mM NaCl; provided by manufacturer. | |||||||
| cThe final ratio of DNA to Nanofectin should be 1:3.2. | |||||||
