Research|Viral Core | Services

Custom lenti viral packaging service

We use the third-generation lentivirus packaging system. Our lentivirus is pseudotyped with VSV-G. We titer lenti preps with Lenti-X™ qRT-PCR Titration Kit (TAKARA). Please provide at least 100 μg plasmid DNA for a lenti prep. Recommended DNA concentration is 0.5ug/ul~2ug/ul. DNA can be dissolved in sterile water.

For lenti vectors with inserts < 7kb between two LTRs, we use ultracentrifugation to concentrate the virus. Final product is in 0.001%F68+DPBS. Typical yield is 150ul~200ul concentrated virus with genomic titer between e+11gc/ml~e+13gc/ml. Virus is good for both in-vivo and in-vitro applications.

For lenti vectors with large inserts (7~10kb), we use lenti-X concentrator (TAKARA) to concentrate the virus. Yield is 800ul~900ul virus with genomic titer between e+8gc/ml~e+10gc/ml. Virus is only good for in-vitro applications.

As lenti viruses are not stable at 4 degree, we aliquot our lenti preps and store at -80 degree till pickup/shipment.

Location of Service:3 Blackfan Circle, CLS13060

Custom AAV packaging service

We purify AAVs using iodixanol gradients and ultracentrifugation. We can package AAV vectors into serotypes 1, 2, 5, 6, 7, 8, 9, rh10, DJ, Anc80, SHH10, retro, 7m8, PHP.S, PHP.b, and PHP.eb. We titer AAVs with Sybr-Green based qPCR assays. Purified AAV is further filter sterilized by a 0.22um filter and is good for both in-vivo and in-vitro applications. Final AAV product is in 0.001%F68+DPBS.

We offer two scales of AAV production. For a regular scale AAV custom prep, typical yield is 250ul~400ul purified virus with genomic titer between e+12gc/ml~e+14gc/ml. Please provide at least 150ug verified plasmid DNA with DNA concentration between 0.5ug/ul~2ug/ul.
For a half scale AAV custom prep, typical yield is 100~150ul purified virus with genomic titer between e+12gc/ml~e+14gc/ml. Please provide at least 80ug verified plasmid DNA with DNA concentration between 0.5ug/ul~2ug/ul.

请确认AAV两个也是我之间的插入s less than 4.7kb. As ITRs might be lost during plasmid amplification, please make sure always digest your plasmid with SmaI after plasmid prep to confirm the ITRs are intact. You are supposed to see a 2.6~3kb backbone band.

As AAVs are stable at 4 degree, we keep fresh AAV preps at 4 degree till pickup/shipment. For long term storage, please make aliquots and store at -80 degree.

Location of Service:3 Blackfan Circle, CLS13060

Off-the-Shelf Premade AAV

We provide pre-made AAVs with different serotypes for AAV-GFP and AAV-Cre. Please contact the Core for details.

Location of Service:3 Blackfan Circle, CLS13060

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How to submit your DNA

Local labs could drop off your DNA into our drop off/pick up fridge. Please label your tube with the plasmid name and your name. The fridge is located in CLS building 13th floor, room 13060. It is next to bench CLS13060.13 and room CLS13065.

Please use the following address if you choose to ship your DNA samples:

Boston Children's Hospital Viral Core
Att: Yiming Zhang
3 Blackfan Circle, CLS13060
Boston, MA, 02115

How to submit your iLab orders

Please submit your orders using theBCH ilab site.

1. If you don’t have an account with ilab, please complete the registration first. Complete the registration form on the sign-up page. The light blue register button is located at the top right corner of the sign-up page. Receive a Welcome Email from iLab (typically within one business day) with login credentials.
2. Go to “Core facilities,” “Viral Core,” “request services,” and place order.

Pricing

If you are a current user, please login to VPN toaccess pricing. If you are a new user, please contact the Core directly to request services and specific pricing information.