TABLE 1.
Troubleshooting table.
| PROBLEM | POSSIBLE REASON | SOLUTION |
|---|---|---|
| Microcarriers agglomerate after coating with DNA. |
DNA concentration is too high. | Reduce the DNA concentration. |
| DNA is impure. | Check the purity of the DNA by running a gel. | |
| Gold does not spread evenly in the tubing once the tube-preparation station is switched on. |
Old PVP solution. | Make a new stock of PVP in ethanol; use a fresh bottle of ethanol. |
| Insufficiently dry tube before loading. | Flush tubing with nitrogen for 5–10 min prior to loading with DNA/gold microcarrier solution. |
|
| Poor or no expression. | Poor cell or tissue health. | Use young healthy cells or tissue. |
| DNA concentration too low. | Increase the DNA concentration. | |
| DNA is impure. | Check the DNA purity by running a gel. | |
| Spermidine needs replacing. | Make a new solution of spermidine. | |
| Contamination of cell cultures. | Poor culture conditions. | Improve culture conditions. |
| Gene gun is contaminated. | Wash the end of the barrel with 70% (vol/vol) ethanol and change the nylon mesh. |
|
| Cell death. | Poor cell or tissue health. | Use young healthy cells or tissue. |
| Gas pressure too high. | Reduce gas pressure. | |
| Microcarriers too large. | Use a smaller size. |