Gene Transfer, Targeting and Therapeutics
Viral Vector Core

Services

• Production of custom lentiviral, retroviral, AAV, G-deleted rabies, and adenoviral vectors
• Production and distribution of stock lentiviral, retroviral, AAV, G-deleted rabies, and adenoviral vectors expressing reporter transgenes
• Consultation on experimental design, development and use of custom viral vectors
• Production and distribution of AAV serotype kits useful for pilot studies

What to provide to the core:

1. 200ug (preferably >1ug/ul) of your lentiviral or AAV plasmid. If large scale AAV production is requested, 500ug of AAV plasmid is required. DNA should have been purified using an endotoxin-free protocol (e.g. Endo-free maxi/mega/giga plasmid purification kits or equivalent). We do not accept miniprep purified DNA.
2. Plasmid DNA should be checked for purity and have an A260/280 of >1.8.
3. AAV ITR plasmids should be checked for recombination by digestion with SmaI or XmaI. Each ITR contains two SmaI/XmaI sites; digestion will cut out your insert. Excessive amounts of linearized full-length plasmid indicate recombination has occurred.
4. Lentiviral and retroviral transfer plasmids should be confirmed by enzyme digest or sequencing.
5. Please provide a gel image of your SmaI or XmaI digested AAV2 ITR plasmid, or your lentiviral transfer plasmid with the insert excised.
6. To avoid recombination we recommend transforming and growing your lentiviral and AAV transfer plasmids in recombination deficient cells such as STBL3 @ 30°C IN 2XYT broth for no more that 16 hrs.
7. Vector map and Sequence file, if available.
8. Information about your gene of interest or insert any special requirements for handling: toxic, oncogenic, pro-apoptotic, etc.