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pAAV-GFAP104-mCherry
(Plasmid #58909)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 58909 Standard format: Plasmid sent in bacteria as agar stab 1 $65
AAV5 58909-AAV5 Virus (100 µL at titer ≥ 1×10¹³ vg/mL) and Plasmid. More Information
$380

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    AAV
  • Backbone size w/o insert (bp) 4895
  • Total vector size (bp) 5606
  • Vector type
    Mammalian Expression, AAV

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    NEB Stable
  • Growth instructions
    Stbl3 at 30C (use carbenicillin if using Stbl3) OR DH5a at 37C
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    mCherry
  • Species
    Synthetic
  • Insert Size (bp)
    711
  • Promoter GFAP

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BamHI (not destroyed)
  • 3′ cloning site EcoRI (not destroyed)
  • 5′ sequencing primer ggagagctctccccatag
  • 3′ sequencing primer cagcgtatccacatagcg
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

The depositing lab has sequenced the 5' ITR, promoter, and fluorophore. Multiple digestions were done to verify the vector structure. The construct was tested in vitro and the virus was tested both in vitro and in vivo.

Information for AAV5 (Catalog # 58909-AAV5) ( Back to top )

Purpose

Ready-to-use AAV5 particles produced from pAAV-GFAP104-mCherry (#58909). In addition to the viral particles, you will also receive purified pAAV-GFAP104-mCherry plasmid DNA.

GFAP104-driven mCherry expression control. These AAV preparations are suitable purity for injection into animals.

Delivery

  • 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, AAV5 cap gene
  • Buffer PBS + 0.001% Pluronic F-68 + 200 mM NaCl
  • Serotype AAV5
  • Purification Iodixanol gradient ultracentrifugation
  • Reporter Gene mCherry

Biosafety

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

Resource Information

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). 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.

How to cite this plasmid ( Back to top)

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-GFAP104-mCherry was a gift from Edward Boyden (Addgene plasmid # 58909 ; http://n2t.net/addgene:58909 ; RRID:Addgene_58909)

    For viral preps, please replace (Addgene plasmid # 58909) in the above sentence with: (Addgene viral prep # 58909-AAV5)

  • For your References section:

    Optogenetic astrocyte activation modulates response selectivity of visual cortex neurons in vivo. Perea G, Yang A, Boyden ES, Sur M. Nat Commun. 2014;5:3262. doi: 10.1038/ncomms4262. 10.1038/ncomms4262 PubMed 24500276