Schedule of Charges
Effective August 16, 2011
Please note: Core services and prices are subject to change. Pricing is reviewed regularly by the Core Oversight Committee. Washington University neuroscience investigators and Hope Center member labs may be eligible for credits for services through the P30 Blueprint Core and Hope Center respectively. An additional 25% surcharge will be applied to orders from academic and non-profit institutions outside of Washington University.
Viral Vectors Core price structure: TBD= to be determined by agreement between core and user. The prices in the list are before subsidy for eligible internal users and do not include a 25% surcharge for external academic users.
|4a||PLASMID PREP (for making virus in the Core)||Each||$200.00|
|5||LENTIVIRUS PREP: Large Scale||Each||$1,500.00|
|5a||LENTIVIRUS PREP: Large Scale (order 2 or more)||Each||$1,200.00|
|6||LENTIVIRUS PREP: Small Scale (10 ml crude supernatant)||Each||$250.00|
|7||LENTIVIRUS PREP: Small Scale (purified and titered)||Each||$750.00|
|7a||LENTIVIRUS PREP: Small Scale (purified and titered – order 2 or more)||Each||$600.00|
|8||VIRAL TITER: FACS/qPCR/Cell count (included in viral preps)||Each||$300.00|
|10||Use of core hoods||Hour||$60.00|
|12||LENTI ALIQUOT sampler (choose 3 among PGK, CMV, ubiq, Synapsin, GFAP promoter-GFP virus, small aliquots)||Each||$60.00|
|13||AAV ALIQUOT sampler (AAV1, AAV2, AAV5, small aliquots)||Each||$60.00|
|17||Consultation I||1/2 Hour||$20.00|
|18||Consultation II||1/2 Hour||$40|
|19||In Lab Training||1 Hour||$60.00|
|20||AAV PREP: Large scale||Each||$3000.00|
|20a||AAV PREP: Large scale (order 2 or more)||Each||$2400.00|
Note: If the titer of a viral vector prep is below the expected range, Viral Vector Core will identify the cause of the problem and make another attempt to produce the requested viral vector. If low titer was likely due to mistakes during the vector production and purification, the core will make a new prep without charge. If low titer is caused by a larger than optimal insert, mutation, deletion, or impurity of transfer plasmid supplied by the user, the PI will be billed. In rare cases, it may not be possible to make a high titer prep with specific inserts. The core will work with investigators to maximize titers in this situation and will keep users informed of charges for multiple preps.
Detailed description of services
- Subcloning: For labs that do not have the expertise to create their own constructs, the viral core will insert a gene of interest into lentiviral or AAV transfer vector. User provides source DNA with detailed map and sequences of the plasmid (hard copy OK but electronic preferred).
Simple subcloning: Gene of interest can be inserted using restriction enzyme digestion and ligation.
More complex: restriction site needs to be added to gene of interest, protein tags added, mutations introduced, etc.
Custom/Develop new vector: For example, add new promoter to viral vectors. Available promoters in lentivirus now are CMV, PGK, MPB, MND, GFAP and ubiquitin.
- Plasmid preparation: Core will prepare a “Mega” prep of plasmid DNA suitable for use in lentiviral transfection. User provides small amount of DNA. Typical yields are at least 500 μg DNA (> 1 mg for high copy number plasmids).
- Lentivirus preparation: Core transfects viral transfer vector and generates lentivirus. If user provides transfer vector, user is responsible for insuring that vector is cloned correctly with all required sequences for viral production. Virus is purified by ultracentrifugation, titer (≥5 x10^7, typically 10^8/ml). Large scale prep is 10-20 x 10 cm dishes. Small scale prep is a 6 well culture vessel. Small scale plus purification and titration is 5 x 10 cm dishes. User provides 500 μg of purified transfer plasmid DNA for large scale prep (or core prepares plasmid as above). Titering by FACS or real time PCR is included. Viruses expressing other markers (lacZ, myc, HA tags, proteins for which an antibody is available) can be titered by serial dilution, immunochemistry and cell counting for additional charge. User should provide a protocol for non-standard epitopes.
- Lentivirus aliquot: Aliquots of purified viruses are available. Titers are at least 5 x 10^7/ml. Aliquot size is at least 10 μl.
- AAV vector preparation:User provides 1mg of purified transfer plasmid DNA (or core prepares mega DNA prep at additional charge). NOTE: User is responsible for insuring that AAV vector is cloned correctly. Core may do diagnostic digests to confirm that vectors are correct. Core transfects viral transfer vector and AAV helper plasmids to produce AAV vector with serotype of choice. AAV vector is purified by Iodixanol gradient centrifugation and affinity or ion exchange column. Titer is determined by Dot blot analysis and is typically 10^11 – 10^12 vector genome/ml. Infectious center assay is performed for every batch of vectors for our internal quality control. Transduction efficiency is routinely tested in murine neocortical cultures provided that the construct contains a fluorescence reporter.
- AAV vector aliquot: Aliquots of purified AAV vectors are available. Titers are at least 1 x 10^11/ml. Aliquot size is at least 10 μl.
- Other: Core training in protocols and techniques is billed on a time-based basis.
Use of core supplies and equipment is billed on a cost recovery basis.
Use of core hoods is charged at $60 per hour in half-hour increments.
Consultation: Routine discussions of projects are covered under costs of projects. More extensive consultation and training is billed on a time basis and also on the number of core staff involved.
Biotechnology Building, Room 221
Biotechnology Building, Room 220
We have aliquots of most stock viruses available for immediate use. Preparation of a new virus can take 3-6 weeks, depending on complexity of subcloning, transduction efficiency of constructs, etc.