Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | ||
|---|---|---|---|---|---|---|
| Plasmid | 51085 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
| AAV1 | 51085-AAV1 | Virus (100 µL at titer ≥ 7×10¹² vg/mL)and Plasmid.More Information | Add to Cart | |||
| AAV Retrograde | 51085-AAVrg | Virus (100µL at titer ≥ 7×10¹² vg/mL)and Plasmid.More Information | Add to Cart | |||
This material is available to academics and nonprofits only.
Backbone
- Vector backbonepAAV-hSyn1-WPRE-HGHpA(Search Vector Database)
- Backbone manufacturerModified from Karl Deisseroth
- Backbone sizew/o insert(bp)4590
- Total vector size (bp)6750
- Modifications to backboneAltered cloning sites
- Vector typeMammalian Expression, AAV
Growth in Bacteria
- Bacterial Resistance(s)Ampicillin
- Growth Temperature37°C
- Growth Strain(s)Stbl3
- Copy numberHigh Copy
Gene/Insert
- Gene/Insert nameGCaMP6f
- Alt nameG6f
- Alt nameGCaMP6 fast
- SpeciesSynthetic
- Insert Size (bp)2160
- Promoterhuman Synapsin1
- Tag/ Fusion Protein
- P2A-nls-dTomato (C terminal on insert)
Cloning Information
- Cloning methodRestriction Enzyme
- 5′ cloning siteBamHI(not destroyed)
- 3′ cloning siteAscI(not destroyed)
- 5′ sequencing primerctcagcgctgcctcagtctg
- 3′ sequencing primerCCATACGGGAAGCAATAGCATG (Common Sequencing Primers)
Resource Information
- A portion of this plasmid was derived from a plasmid made byThe GCaMP6f portion was derived from Addgene plasmid #40755 (Douglas Kim, Janelia Farms).
- Terms and Licenses
- UBMTA
- Ancillary Agreement for Plasmids Containing FP Materials
- Takara Bio Limited Use Label License (formerly Clontech)
- Industry Terms
- Not Available to Industry
- Articles Citing this Plasmid
- 6 References
Depositor Comments
The addition of the P2A-nls-dTomato allows for easy identification of GCaMP6 expressing cells exhibiting nuclear red fluorescence that does not significantly overlap with the cytosolic GCaMP signal.The GCaMP and fluorophore are physically uncoupled.
Permissions were obtained from Douglas Kim and the Clontech licensing office for depositing this new plasmid.
Information for AAV1 (Catalog # 51085-AAV1)(Back to top)
Purpose
Ready-to-use AAV1 particles produced from AAV-hSyn1-GCaMP6f-P2A-nls-dTomato (#51085). In addition to the viral particles, you will also receive purified AAV-hSyn1-GCaMP6f-P2A-nls-dTomato plasmid DNA.
GCaMP6f calcium sensor and bicistronic, physically separate nuclear localized dTomato expression under the Synapsin promoter. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 7×10¹² vg/mL
- Pricing$350 USD for preparation of 100 µL virus + $30 USD for plasmid.
- StorageStore at -80℃. Thaw just before use and keep on ice.
- ShipmentViral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
- Packaging Plasmidsencode adenoviral helper sequences and AAV rep gene, AAV1 cap gene
- BufferPBS + 0.001% Pluronic F-68 + 200 mM NaCl
- SerotypeAAV1
- PurificationIodixanol gradient ultracentrifugation
- Reporter Genenuclear dTomato (physically separate, not a fusion protein)
Biosafety
Requestor is responsible for compliance withtheir institution"s biosafety regulations.Lentivirus is generally considered BSL-2. AAV isgenerally considered BSL-1, but may requireBSL-2 handling depending on the insert.Biosafety Guide
Resource Information
- Terms and Licenses
- Ancillary Agreement for Penn Vectors
- Terms of Use for Viral Vectors
- Industry Terms
- 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). Thespecific 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 moreinformation.
Information for AAV Retrograde (Catalog # 51085-AAVrg)(Back to top)
Purpose
Ready-to-use AAV Retrograde particles produced from AAV-hSyn1-GCaMP6f-P2A-nls-dTomato (#51085). In addition to the viral particles, you will also receive purified AAV-hSyn1-GCaMP6f-P2A-nls-dTomato plasmid DNA.
GCaMP6f calcium sensor and bicistronic, physically separate nuclear localized dTomato expression under a human synapsin1 promoter. These AAV were produced with a retrograde serotype, which permits retrograde access to projection neurons.These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 7×10¹² vg/mL
- Pricing$350 USD for preparation of 100 µL virus + $30 USD for plasmid.
- StorageStore at -80℃. Thaw just before use and keep on ice.
- ShipmentViral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
- Packaging Plasmidsencode adenoviral helper sequences and AAV rep gene, AAV retrograde cap gene from rAAV2-retro helper (plasmid #81070)
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV retrograde (AAVrg)
- PurificationIodixanol gradient ultracentrifugation
- Reporter Genenuclear dTomato (physically separate, not a fusion protein)
Biosafety
Requestor is responsible for compliance withtheir institution"s biosafety regulations.Lentivirus is generally considered BSL-2. AAV isgenerally considered BSL-1, but may requireBSL-2 handling depending on the insert.Biosafety Guide
Resource Information
- Terms and Licenses
- Terms of Use for Viral Vectors
- Industry Terms
- 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). Thespecific 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 moreinformation.
Addgene Comments
Retrograde functionality is dependent on high viral titers. Addgene recommends not diluting your AAV preps prior to use.Data submitted about 51085-AAVrg by requesting scientist(s):
- Data 1: Mouse, Brain parenchyma
