|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||105714||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
|AAV8||105714-AAV8||Virus (100 µL at titer ≥ 1×10¹³ vg/mL) and Plasmid.|
This material is available to academics and nonprofits only.
Vector backbonepAAV-Ef1a-fDIO hChR2(H134R)-EYFP
Backbone manufacturerKarl Deisseroth
- Backbone size w/o insert (bp) 5609
- Total vector size (bp) 6857
Vector typeMammalian Expression, AAV ; Flp/Frt
Growth in Bacteria
Growth Strain(s)NEB Stable
Copy numberHigh Copy
Alt nameGCaMP3-K78H T302L R303P D380Y T381R S383T R392G
Alt nameGCaMP3 variant 641
SpeciesR. norvegicus (rat); A. victoria (jellyfish)
Insert Size (bp)1274
- Promoter Ef1a, Human elongation factor-1 alpha
- Cloning method Restriction Enzyme
- 5′ cloning site NheI (not destroyed)
- 3′ cloning site AscI (not destroyed)
- 5′ sequencing primer cCTATTCTTCaAAAGGTATAGGAACTTC
- 3′ sequencing primer tTCTAGAGAATAGGAACT (Common Sequencing Primers)
A portion of this plasmid was derived from a plasmid made byGCaMP6s was synthesized de novo based on sequences published by Douglas Kim et al, Janelia Research Campus in Chen et al, 2013 PMID: 23868258.
Terms and Licenses
- Not Available to Industry
Information for AAV8 (Catalog # 105714-AAV8) ( Back to top )
Ready-to-use AAV8 particles produced from pAAV-Ef1a-fDIO-GCaMP6s (#105714). In addition to the viral particles, you will also receive purified pAAV-Ef1a-fDIO-GCaMP6s plasmid DNA.EF1a-driven, Flp recombinase-dependent expression of GCaMP6s calcium sensor. These AAV preparations are suitable purity for injection into animals
- Volume 100 µL
- Titer ≥ 1×10¹³ 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, AAV8 cap gene
- Buffer PBS + 0.001% Pluronic F-68
- Serotype AAV8
- 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
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 CommentsUsing recombinase-dependent vectors in vivo: FRT sites in fDIO plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Flp-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.01-0.03% of viral vectors in our typical production protocol. This can lead to a small number of cells exhibiting Flp-independent transgene expression in vivo. To address this, we recommend titrating to find the optimal AAV dosage required for Flp-dependent transgene expression and function in vivo. This may include reducing the viral vector dosage in order to reduce the likelihood of Flp-independent 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-Ef1a-fDIO-GCaMP6s was a gift from Rylan Larsen (Addgene plasmid # 105714 ; http://n2t.net/addgene:105714 ; RRID:Addgene_105714)
For viral preps, please replace (Addgene plasmid # 105714) in the above sentence with: (Addgene viral prep # 105714-AAV8)