1. Note about the 2014 updates to our Terms and Conditions
To our customers who have previously utilized our paper-based order form, please note that our Terms and Conditions have changed to reflect our current electronic format. We recommend reading them again before submitting a Service Request. By submitting a Service Request, the User acknowledges that h/she has read and agreed to our Terms and Conditions.
2. Access and Utilization
The Massachusetts General Hospital Vector Core Facility (“the Core”) will be open to investigators affiliated with Partners HealthCare System (PHS), any other academic institutions, and industry. Prices for industrial users must be pre-negotiated with our Director; please click here for contact information. Virus production will be performed in the order that Service Requests are received by the Core.
To submit a Service Request, Users must Register with the Partners Institutional Biosafety Committee (PIBC). All our Service Request forms require a valid PIBC registration number. Labs who have previously registered with the Harvard University Committee on Microbiological Safety (COMS) and have not registered with the PIBC must do so before submitting a Service Request. COMS numbers are no longer acceptable, and absolutely no projects will be initiated without PIBC approval.
Users of the Core will create an Account. After this point, all interactions with the Core will occur through the website or the Core email address: firstname.lastname@example.org
The Core reserves the right to refuse work, in the case of transgenes the Director deems unsafe for production, or when a conflict of interest arises, in which case the User will be directed to other core facilities in the country. Standard hours of operation will be Monday through Friday, 9am to 5pm. The packaging cycle begins each Monday; plasmids received after Monday will be entered into the packaging cycle for the following week. The Core will be closed on all official holidays at Massachusetts General Hospital.
By using our services, you agree to participate in our Plasmid Sharing Program which allows investigators interested in your submitted construct to contact you and request your approval for their use and be packaged by our core.
Billing will take place after completion of service. Electronic invoices will post to the Invoices tab of the User menu on the MGH Vector Core website. PHS-affliated investigators will be asked to provide a PeopleSoft number to be charged for the amount corresponding to the service. For non-PHS investigators and companies, a PO for the amount of the services to be rendered must be provided we begin any work. Once the Core has completed the requested service(s), we will contact the AR Department of the non-PHS investigator or company to collect payment, which must be paid by check.
4. Plasmid Requirements and Specifications
The User requesting the service is required to provide the Core with sterile vector DNA prepared with an endotoxin-free maxi kit. Because of the large volume of plasmids we receive, we request that Users submit their DNA in an Eppendorf tube labeled with the plasmid name provided on the Service Request form (and not a nickname or abbreviation used in their lab). We also request that plasmids be diluted to a concentration of 1 µg/µl, at the following amounts, depending on the Service desired:
|Service requested||Amount of DNA required|
|AAV preparation||400 µg|
|Lentivirus (concentrated preparation)||400 µg|
|Lentivirus (small-scale preparation)||20 µg|
|Retrovirus (concentrated preparation)||400 µg|
Alternatively, the Core is able to produce transfection-quality DNA in our own facilities. To learn more about prepping your plasmid at the MGH Vector Core Facility, see below. Please keep in mind that the quality of your DNA affects quite significantly the final titer of your virus.
If using a plasmid obtained through the the Core, or, if using an MVC whose structural integrity will be modified in the cloning process, the researcher is released from the responsibility of demonstrating its structural and functional integrity. For researchers providing their own plasmids, the Core requires the following information before initiating virus production:
- A clear map of the plasmid showing all coding and non-coding genetic elements present in the vector and the restriction sites used to determine its structural integrity.
- The source of the polyA signal used in AAV vectors. Presently, the MVC uses primers and probes specific for the bovine growth hormone (BGH) polyA signal to determine the AAV vector genome titer. In the absence of this element, the titration method should be discussed prior to initiation of service. For researchers submitting lentiviral constructs without a WPRE, titration will not be possible.
- Users should perform a restriction enzyme digest of their vector and upload a digital image with their Service Request in order to demonstrate its structural integrity.
- Users are to test the functionality of their plasmid by transient transfection in vitro. Users who do not attempt to test the function of their constructs in vitro do so against the advice of the MGH Vector Core Facility, and will not be offered the opportunity to have their virus re-packaged should the first attempt at packaging functional viral particles fail (see below).
Please refer to this Guide for non-MVC Plasmid Construction for more information.
Once requirements 4.1-4.4 have been met, the Core will initiate vector production for Users submitting DNA with non-MVC plasmids.
5. Virus Production and Service Delivery
The MVC will provide Users with electronic Order Status updates on the MVC website for Users to monitor the progress of the their order. The MVC Order Status updates are as follows:
- Project Initiated: The User has submitted a Service Request on the MVC website.
- Plasmid Received: The MVC has received plasmid DNA from the User.
- Packaging in Progress: Virus production has begun.
- Purification in Progress: For AAVs only, this indicates isolation and purification of AAVs.
- Completed: Virus is titered, aliquoted, and ready for shipping or pick-up.
Once a project is completed, the User may contact the MVC to arrange a date for pickup. If the User prefers to have the virus shipped, the MVC will ship the virus using the FedEx account number and shipping address provided on the Account page. Please note that the MVC will ship by FedEx Overnight Express, exclusively, and that the Recipient FedEx account number provided by the User will be billed for the cost of shipping. The MVC will add $25 to the User’s invoice to cover the cost of supplies.
The MVC will make up to two attempts to produce the requested virus. If the viral titer or yield is low, possible explanations include the following:
- Toxicity or deleterious effects on packaging due to encoded transgene(s).
- Mutations or deletions in cis-regulatory elements necessary for packaging, e.g. ITRs, packaging signal.
- Low-quality plasmid DNA.
- Mishaps during the virus production process.
If the cause of low yield is due to 5.4, the MVC will attempt another preparation without charge. (User may be asked to provide more plasmid DNA). Please note that if the cause of low yield is due to 5.1-5.3, the User will be billed for the service.
Users submitting their own, non-MVC plasmid who did not attempt to test the activity of their construct before will not be offered a re-package in the event that their viruses have no measurable biological activity, and they will be billed for the service. Again, Users are to test their constructs in vitro before submitting them for packaging.
6. Plasmid production
The Core offers Users the option of preparing sterile, endotoxin-free DNA for transfection. This service is only available to Users who will submit a Service Request for virus preparation with us; we will not prepare DNA for routine laboratory use.
7. Virus aliquots
8. Publications and Citations
The Core must keep records for the NIH of PIs who have published papers using our viruses. Therefore, by submitting a Service Request, the User the agrees to:
- Inform the Core of any papers he/she publishes using a virus derived from the Core.
- Acknowledge the Core in his/her paper, and
For more information, please refer to our Publications and Citations page
It is the responsibility of the User and his/her PI to make sure that use of viruses is in compliance with his/her Institution’s biosafety regulatory entity. This may include institutional registration of the virus, and, for in vivo applications, Institutional Animal Care and Use Committee (IACUC) approval of any animal protocols utilizing the virus before initiating experiments. No viruses received from the Core are suitable for use in humans. The MGH Vector Core Facility, the Massachusetts General Hospital, and Partners HealthCare are not responsible for the virus once it is released to the the User and his/her PI.
Utilization of the MGH Vector Core Facility does not entitle a User to any of the following: reagents (polyebrene, cell lines, antibodies, luciferase substrates) technician time, lab space (e.g., benches and cell culture hoods) or equipment (e.g., luminometer, fluorescent microscope, RT-PCR machines).