PurposeExpresses GCaMP6f under control of eukaryotic Ef1a promoter after Flp-dependent recombination.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||128315||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
Limited Stock Available, 2 units left
Virus (100 µL at titer ≥ 1×10¹³ vg/mL) and Plasmid.
This material is available to academics and nonprofits only.
Backbone manufacturerAddgene # 55641
- Backbone size w/o insert (bp) 6323
- Total vector size (bp) 6965
Modifications to backboneEYFP cDNA removed by AscI-NheI double digest
Vector typeMammalian Expression, AAV
Growth in Bacteria
Bacterial Resistance(s)Ampicillin, 100 μg/mL
Growth Strain(s)NEB Stable
Copy numberHigh Copy
Insert Size (bp)1371
- Promoter Ef1a
/ Fusion Protein
- His Tag (N terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site AscI (destroyed during cloning)
- 3′ cloning site NheI (destroyed during cloning)
- 5′ sequencing primer CACCCACACAAAGGAAAAGGGCC
- 3′ sequencing primer GCAATAGCATGATACAAAGG (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
Information for AAV1 (Catalog # 128315-AAV1) ( Back to top )
Ready-to-use AAV1 particles produced from pAAV-Ef1a-fDIO-GCaMP6f (#128315). In addition to the viral particles, you will also receive purified pAAV-Ef1a-fDIO-GCaMP6f plasmid DNA.EF1a-driven, Flp-dependent expression of calcium sensor GCaMP6f. 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, AAV1 cap gene
- Buffer PBS + 0.001% Pluronic F-68
- Serotype AAV1
- 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.
Using 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 0.1-0.8% of viral particles in our typical production protocol. This can lead to a small number of cells exhibiting Flp-independent transgene expression in vivo. To address this, it is necessary to optimize the injection volume and viral titer to find the optimal AAV dosage required for Flp-dependent transgene expression and function in vivo. This may include reducing the viral particle 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-GCaMP6f was a gift from Patricia Jensen (Addgene plasmid # 128315 ; http://n2t.net/addgene:128315 ; RRID:Addgene_128315)
For viral preps, please replace (Addgene plasmid # 128315) in the above sentence with: (Addgene viral prep # 128315-AAV1)
For your References section:Natural locus coeruleus dynamics during feeding. Sciolino NR, Hsiang M, Mazzone CM, Wilson LR, Plummer NW, Amin J, Smith KG, McGee CA, Fry SA, Yang CX, Powell JM, Bruchas MR, Kravitz AV, Cushman JD, Krashes MJ, Cui G, Jensen P. Sci Adv. 2022 Aug 19;8(33):eabn9134. doi: 10.1126/sciadv.abn9134. Epub 2022 Aug 19. 10.1126/sciadv.abn9134 PubMed 35984878