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Addgene/pAAV-CAG-Flex-mRuby2-GSG-P2A-GCaMP6f-WPRE-pA/1unit/68719-AAV1

Ordering

Item	Catalog #	Description	Quantity	Price (USD)
Plasmid	68719	Standard format: Plasmid sent in bacteria as agar stab	1	$75	Add to Cart
AAV1	68719-AAV1	Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information		$380	Add to Cart

This material is available to academics and nonprofits only.

Backbone

Vector backbone
pAAV
(Search Vector Database)
Total vector size (bp)7112
Vector type
AAV

Growth in Bacteria

Bacterial Resistance(s)
Ampicillin
Growth Temperature
37°C
Growth Strain(s)
Stbl3
Growth instructions
Due to ease of recombination, AAV and lentivirus vectors should be amplified in a recombination deficient bacteria strain such as Invitrogen's OneShot Stbl3 cells. Check for integrety of ITR sites with SmaI digest.
Copy number
Low Copy

Gene/Insert

Gene/Insert name
mRuby2-P2A-GCaMP6f
Species
R. norvegicus (rat), G. gallus (chicken); A. victoria (jellyfish)
Insert Size (bp)
2124
PromoterCAG-FLEX

Cloning Information

Cloning methodUnknown
5′ sequencing primeractcagcgctgcctcagtct
3′ sequencing primergtttgtacaaatgatgacagcgaag

Resource Information

Terms and Licenses
- UBMTA
- Ancillary Agreement for Plasmids Containing FP Materials
- genOway Notice of RIghts
Industry Terms
- Not Available to Industry

Depositor Comments

The discrepancies between the full sequence and Addgene's QC sequence should not have any functional consequence.

mRuby2:Article: Improving FRET dynamic range with bright green and red fluorescent proteins. Lam et al (Nat Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171. PubMed)Addgene Plasmid 40260

GCaMP6f:Article: Ultrasensitive fluorescent proteins for imaging neuronal activity. Chen et al (Nature. 2013 Jul 18;499(7458):295-300. doi: 10.1038/nature12354. PubMed)Addgene Plasmid 40755

Information for AAV1 (Catalog # 68719-AAV1)(Back to top)

Purpose

Ready-to-use AAV1 particles produced from pAAV-CAG-Flex-mRuby2-GSG-P2A-GCaMP6f-WPRE-pA (#68719). In addition to the viral particles, you will also receive purified pAAV-CAG-Flex-mRuby2-GSG-P2A-GCaMP6f-WPRE-pA plasmid DNA.

CAG-driven, Cre-dependent GCaMP6f calcium sensor with bicistronic mRuby2. These AAV preparations are suitable purity for injection into animals.

Delivery

Volume100 µL
Titer≥ 1×10¹³ 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
SerotypeAAV1
PurificationIodixanol gradient ultracentrifugation
Reporter GenemRuby2 (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
- Ancillary Agreement for Penn Vectors
- Terms of Use for Viral Vectors
Industry Terms
- Not Available to Industry

Viral Quality Control

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

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.

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Addgene/pAAV-CAG-Flex-mRuby2-GSG-P2A-GCaMP6f-WPRE-pA/1unit/68719-AAV1

Ordering

Backbone

Growth in Bacteria

Gene/Insert

Cloning Information

Resource Information

Depositor Comments

Information for AAV1 (Catalog # 68719-AAV1)(Back to top)

Purpose

Delivery

Viral Production & Use

Biosafety

Resource Information

Viral Quality Control

Addgene Comments