PurposeTo generate Adeno-Associated viruses for expression of dLight1.1 under CAG promoter
Full plasmid sequence is not available for this item.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||111067||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
|AAV5||111067-AAV5||Virus (100 µL at titer ≥ 7×10¹² vg/mL) and Plasmid.|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 5020
- Total vector size (bp) 7063
Growth in Bacteria
Growth Strain(s)NEB Stable
Copy numberHigh Copy
Insert Size (bp)2043
/ Fusion Protein
- Flag tag (N terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site BaMHI (not destroyed)
- 3′ cloning site HindIII (not destroyed)
- 5′ sequencing primer GCT CTA GAG CCT CTG CTA ACC ATG TTC
- 3′ sequencing primer cgggaagcaatagcatgatacaaaggc (Common Sequencing Primers)
Addgene NGS identified chicken beta-actin promoter, but not the CMV enhancer and chimeric intron often associated with CAG promoters. Please see the associated diagnostic digest for plasmid verification.
Information for AAV5 (Catalog # 111067-AAV5) ( Back to top )
Ready-to-use AAV5 particles produced from pAAV-CAG-dLight1.1 (#111067). In addition to the viral particles, you will also receive purified pAAV-CAG-dLight1.1 plasmid DNA.CAG-driven expression of dLight1.1 dopamine sensor. This sensor is recommended for photometry in vivo. These AAV preparations are suitable purity for injection into animals.
- Volume 100 µL
- Titer ≥ 7×10¹² vg/mL
- Pricing $350 USD for preparation of 100 µL virus + $30 USD for plasmid.
- Storage Store at -80℃. Thaw just before use and keep on ice.
- Shipment Viral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
- Packaging Plasmids encode adenoviral helper sequences and AAV rep gene, AAV5 cap gene
- Buffer PBS + 0.001% Pluronic F-68 + 200 mM NaCl
- Serotype AAV5
- Purification Iodixanol gradient ultracentrifugation
Requestor is responsible for compliance with their institution's biosafety regulations. Lentivirus is generally considered BSL-2. AAV is generally considered BSL-1, but may require BSL-2 handling depending on the insert. Biosafety Guide
Terms and Licenses
- Not Available to Industry
Viral Quality Control
- Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC) (see a list of our QC assays). The specific QC assays performed varies for each viral lot. To learn which specific QC assays were performed on your lot, please contact us.
- Titer: the exact titer of your sample will be reported on the tube. The titer you see listed on this page is the guaranteed minimum titer. See how titers are measured.
Visit our viral production page for more information.
Addgene CommentsPlease note titers for 111067-AAV5 tend to be low and therefore a higher volume of the viral preparation may be required to see expression than for other AAV5 vectors. Users should titrate the virus to determine the optimal dose for expression.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pAAV-CAG-dLight1.1 was a gift from Lin Tian (Addgene plasmid # 111067 ; http://n2t.net/addgene:111067 ; RRID:Addgene_111067)
For viral preps, please replace (Addgene plasmid # 111067) in the above sentence with: (Addgene viral prep # 111067-AAV5)
For your References section:Ultrafast neuronal imaging of dopamine dynamics with designed genetically encoded sensors. Patriarchi T, Cho JR, Merten K, Howe MW, Marley A, Xiong WH, Folk RW, Broussard GJ, Liang R, Jang MJ, Zhong H, Dombeck D, von Zastrow M, Nimmerjahn A, Gradinaru V, Williams JT, Tian L. Science. 2018 Jun 29;360(6396). pii: science.aat4422. doi: 10.1126/science.aat4422. Epub 2018 May 31. 10.1126/science.aat4422 PubMed 29853555