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INFORMATION FOR

High Titer Virus Services

Overview

Retroviruses and its special subtype, lentiviruses, are essential tools to deliver genetic materials into mammalian hematopoietic cells. However, high titer viruses are required in many hematology applications, yet researchers are often troubled with the low titer of viruses that cannot be effectively used in their research applications. Due to this reason, YCCEH provides multiple services on high titer virus to the hematology community.

Services

Advisory Service and Protocol Service:

YCCEH provides free service to advice experimental planning using viruses, including, but not limited to, choices of retro vs lenti virus, choices of vectors and safety considerations.

In addition, YCCEH provides for free proven protocols for high titer virus production upon request.

Production of High Titer Viruses:

YCCEH provides services to make viruses for researchers. The standard service involves generation of virus in 10 cm cell culture plates, yielding >15 ml of virus. Viruses will be packaged in 293T cells, using standard amphitropic packaging plasmids (i.e. with VSVG as envelop protein). Viruses will be filtered through 0.45 micron filter before delivering to researchers to remove any contaminating packaging cells.

Viral construct needs to be provided to YCCEH, with full disclosure of genetic composition due to safety considerations. Plasmid needs to be high quality midi or maxi prep product.

Cost: $75 per virus for the standard service. $75 additional for plasmid transformation and midi prep to be performed by YCCEH.

Additional services: see Concentrating Virus and Measuring Virus Titer

Concentrating Viruses:

YCCEH provides services to concentrate viruses for researchers. This service is an add-on to the High Titer Virus Production service. We currently do not provide this service as a stand-alone service due to inability to guarantee viral performance.

Concentration step will be performed using high-speed centrifugation. We routinely achieve >20 fold enrichment of virus activity through this concentration step.

Cost: $50 for the first virus. $30 for each additional virus on the same order.

Measuring Viral Titer:

YCCEH provides services to measure virus titer for researchers. This service is an add-on to the High Titer Virus Production service or the Concentrating Virus service.

Viral titer will only be measured if there is a constitutive or doxycycline inducible fluorescence protein marker, or there is a constitutive or doxycycline inducible antibiotics selection marker in the viral construct.

Viral titer will be measured to reflect activity on BaF3 or K562 cells, or both. YCCEH will return the levels of transduction using 3 different viral amounts.

Cost: $60 for the first virus with fluorescent protein marker. $30 for each additional virus on the same order.

Cost: $50 for the first virus with fluorescent protein marker. $25 for each additional virus on the same order.

Contact

Please contact Core A, Jun Lu, at jun.lu@yale.edu.