Table 3.

Troubleshooting Guide for Low-Yield Rabies dG Viruses

Possible Source	Troubleshoot
Packaging plasmids and the transfer vector	Check plasmid integrity with restriction enzyme digestion and/or sequencing prior to use.
Length of insert	Rabies genome in transfer vector should not exceed 8.5 Kb.
Pelleted virus	Remove supernatant from ultracentrifuge tubes carefully to maintain an intact pellet.
Insert	Check if insert is toxic to cells by monitoring transfected packaging cells.
Transgene expression	Check Transgene expression before harvesting supernatant. A longer period of time or larger volumes for infection (higher MOI) may be necessary before harvesting supernatant.