Production of custom lentiviral, retroviral, rAAV, G-deleted rabies, and adenoviral vectors specific for individual investigators needs
Production and distribution of stock lentiviral, retrovial, rAAV, G-deleted rabies, and adenoviral vectors expressing reporter transgenes
Consultation on experimental design, development and use of custom viral vectors, and production of client specific viral transfer plasmids
Production and distribution of rAAV2 serotype kits containing a range of cross-packaged vectors – Useful for pilot studies prior to custom vector production
Distribution of cloning vectors and relevant MTA information to investigators prior to vector design and production
Distribution of validated and titrated iPS reprogramming vector kits
What to provide to the core:
200ug (preferably >1ug/ul) of your lentiviral or rAAV plasmid. If large scale rAAV production is requested, 500ug of rAAV 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.
Plasmid DNA should be checked for purity and have an A260/280 of >1.8.
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.
Lentiviral and retroviral transfer plasmids should be digested with appropriate restriction enzymes to confirm correct insert size.
Please provide a gel image of your SmaI or XmaI digested AAV2 ITR plasmid, or your lentiviral transfer plasmid with the insert excised.
To avoid recombination we recommend transforming and growing your lentiviral and rAAV transfer plasmids in recombination deficient cells such as STBL3 @ 30°C IN 2XYT broth for no more that 16 hrs with shaking.
Vector map and Sequence file – we encourage you to submit your plasmids to our vector library. We will be happy to work with you on establishing MTA agreements for your reagents.
Information about your gene of interest or insert and any special requirements for handling e.g. toxicity, oncogenic, pro-apoptotic, etc.