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The Molecular Cardiology Research Institute/GRASP Adenoviral Vector Core Facility (AVCF), located on the 14th floor of the Tupper Building provides Tufts Medical Center and Tufts University School of Medicine investigators with comprehensive service in the design, creation, and analysis of adenoviral vectors. Serban Georgescu, M.D., Director of the AVCF and Doug Jefferson, Ph.D., of the Cell Culture Core assists with the virus amplification.
The AVCF creates a new recombinant adenovirus for the investigator. Typically, this adenovirus expresses a protein of the investigator's choosing. When the service is complete, we shall supply the investigator with a purified recombinant adenovirus. The anticipated project duration, including amplification and purification of the adenovirus, is 8-10 weeks.
To initiate the service, the investigator must first complete the application and e-mail it to the Administrator. AVCF will then supply an adenovirus shuttle plasmid to the investigator, who will subclone the transgene of interest into this plasmid. (The investigator is responsible for screening the construct to confirm that the correct construct has been made.) The investigator will then return the adenovirus shuttle plasmid to the AVCF, whose staff will then make adenovirus. The newly created adenoviral vector is grown and purified at a large scale.
Investigators who have an adenovirus from an outside source and who need only the amplification and purification services should complete sections II & III of the application and submit it to the Administrator.
Virus Production (including amplification):
Tufts-MC/Tufts investigators: $1,550
Virus Amplification Only:
Tufts-MC/Tufts investigators: $750
CONSIDERATIONS FOR CLONING
1.) Because PmeI and PacI sites are designed to linearize the final construct for transformation and transfection, please avoid using these sites in your inserts.
2.) Please avoid cloning elements that are present more than once in the vector (e.g., CMV promoters) in head-to-head orientations.
3.) Please note that all the shuttle vectors confer resistance to Kanamycin (not ampicilin).
PROTOCOL REQUIREMENTS
Recombinant DNA Protocol
It is the responsibility of each investigator to have Institutional Biosafety Committee approval for the use of rDNA on file with the Tufts University and Tufts Medical Center IBC. The registration form can be downloaded at http://publicsafety.tufts.edu/ehs/?pid=76.
Animal Protocol
It is also the responsibility of each investigator to have an approved animal protocol on file at their institution if their research involves animal models. This protocol must be obtained prior to the submission of an application.
Note: The lab providing this service is not certified by GLP Standards.
Helpful Links
AdEasy System Home Page
AdEasy System Protocol
Vector Maps & Sequences
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