Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | ||
|---|---|---|---|---|---|---|
| Plasmid | 83894 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
| AAV Retrograde | 83894-AAVrg | Virus (100 µL 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(Search Vector Database)
- Vector typeMammalian Expression, AAV, Cre/Lox
Growth in Bacteria
- Bacterial Resistance(s)Ampicillin
- Growth Temperature37°C
- Growth Strain(s)DH5alpha
- Copy numberHigh Copy
Gene/Insert
- Gene/Insert nametdTomato
- SpeciesSynthetic
- PromoterhDlx
Cloning Information
- Cloning methodGibson Cloning
- 5′ sequencing primerchim-int-F (5'-TCTTACTGACATCCACTTTGCC-3')
- 3′ sequencing primerWPRE-R (Common Sequencing Primers)
Resource Information
- Supplemental Documents
- Snapgene Full Sequence
- Terms and Licenses
- UBMTA
- genOway Notice of RIghts
- Takara Bio Limited Use Label License (formerly Clontech)
- Industry Terms
- Not Available to Industry
Information for AAV Retrograde (Catalog # 83894-AAVrg)(Back to top)
Purpose
Ready-to-use AAV Retrograde particles produced from pAAV-hDlx-Flex-dTomato-Fishell_7 (#83894). In addition to the viral particles, you will also receive purified pAAV-hDlx-Flex-dTomato-Fishell_7 plasmid DNA.
mDlx-driven, Cre-dependent expression of dTomato. 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 + 200 mM NaCl
- SerotypeAAV retrograde (AAVrg)
- PurificationIodixanol gradient ultracentrifugation
- Reporter GenedTomato (Cre-dependent)
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.Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-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 Cre-independent transgene expression in vivo. To address this, we recommend titrating to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral vector dosage in order to reduce the likelihood of Cre-independent expression.
