You have just received your virus from Addgene. Now what?
Addgene recommends the following steps to ensure proper handling and storage of your virus for future use. If there is a problem, please contact Addgene within 60 days of receiving your virus. If your virus did not arrive on dry ice, please contact Addgene immediately.
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Recombinant virus, while replication deficient, can infect a broad range of mammalian cell types. Please consult your institutional safety office for guidance on safe handling.
Storage: Store at -80°C
Lentivirus and AAV should be stored immediately at -80°C. For long-term storage, store the virus in the back of the freezer where it is less likely to be influenced by temperature changes associated with opening the freezer door.
Addgene does not recommend subjecting your viral preparation to multiple freeze-thaw cycles as this can reduce the titer. However, if you plan to use the virus for multiple experiments, thaw the viral preparation, aliquot the desired volume, and snap-freeze in liquid nitrogen or a dry ice/ethanol bath before storing at -80°C.
Lentivirus stability at -80°C varies depending on the media composition and the virus identity. Once you receive your virus, Addgene recommends using it within one year.
Use: Thawing and Working with your Virus
Lentiviral stocks should be thawed immediately before use and then kept on ice. While Addgene provides titer values for each lot of virus, transduction efficiency will vary among cell lines. Therefore, Addgene recommends performing transductions with serial dilutions of your viral stock.
Helpful Links and Protocols:
- Lentiviral Guide
- Protocol: Colony Formation Titering Assay for Lentivirus
- Protocol: Generating Stable Cell Lines with Lentivirus
- Complete List of Viral Protocols
- Blog: Viral Vector Posts
AAV stocks should be thawed immediately before use on ice or at room temperature and then kept on ice. AAV viral tropism is determined by many factors and will vary between cell lines or host tissues. For in vitro work, a dose-response/serial dilution study in your cell line(s) of interest is best. For in vivo work, review the literature.
Helpful Links and Protocols:
Learn more about Addgene's procedures for viral production and quality control here. Before beginning your experiment, we encourage you to refer to the primary publication associated with the transfer plasmid. Please use the search bar at the top of the page to search for the catalog number of your viral service material for additional information. If you believe that your viral stock is incorrect, please notify Addgene within 60 days.
Citations in Future Publications
Viruses were prepared using plasmids created by your colleagues. Please acknowledge the Principal Investigator, cite the article listed on the plasmid datasheet, and include the Addgene plasmid number in future publications. Be sure to include citation information for both the viral vector and any packaging plasmids used in the production of your virus.
Thank you for requesting viral service from Addgene. If you have questions or would like to speak with an Addgene scientist, contact us at +1-617-225-9000 or email [email protected].