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pACAGW-ChR2-Venus-AAV
(Plasmid #20071)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 20071 Plasmid sent as bacteria in agar stab 1 $65
AAV1 20071-AAV1 Virus (100 µL at titer ≥ 1×10¹³ vg/mL)
and Plasmid. More Information
$380

This material is available to academics and nonprofits only.

Backbone

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    Stbl3
  • Growth instructions
    Grow on Stbl2 cells
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    Channelrhodopsin-2
  • Alt name
    ChR2
  • Alt name
    Channelrhodopsin
  • Species
    Chlamydomonas reinhardtii
  • Insert Size (bp)
    3088
  • Entrez Gene
    COP4 (a.k.a. CHLREDRAFT_182032)
  • Tag / Fusion Protein
    • Venus (C terminal on insert)

Cloning Information

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

Resource Information

Information for AAV1 (Catalog # 20071-AAV1) ( Back to top )

Purpose

Ready-to-use AAV1 particles produced from pACAGW-ChR2-Venus-AAV (#20071). In addition to the viral particles, you will also receive purified pACAGW-ChR2-Venus-AAV plasmid DNA.

CAG-driven, channelrhodopsin fused to Venus for optogenetic activation. 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, AAV1 cap gene
  • Buffer PBS + 0.001% Pluronic F-68
  • Serotype AAV1
  • Purification Iodixanol gradient ultracentrifugation
  • Reporter Gene Venus

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

Viral Quality Control

Titering Method:
  • Real-time qPCR or digital droplet PCR: The number of genome copies in viral preparations was measured by real-time qPCR or by digital droplet PCR. Titering on these preparations was performed by the University of Pennsylvania Vector Core. Read Addgene's AAV Titration by qPCR protocol here.
Notes:
  • Purity of viral preparation: Viral preparations were subjected to polyacrylamide gel electrophoresis (PAGE) followed by SYPRO Red staining and the molecular weight and relative intensity of the viral capsid proteins was analyzed. The abundance of viral capsid proteins as a fraction of total protein present in the sample was used to determine purity of the AAV preparation.

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:

    pACAGW-ChR2-Venus-AAV was a gift from Karel Svoboda (Addgene plasmid # 20071)

    For viral preps, please replace (Addgene plasmid # 20071) in the above sentence with: (Addgene viral prep # 20071-AAV1)

  • For your References section:

    The subcellular organization of neocortical excitatory connections. Leopoldo Petreanu, Tianyi Mao, Scott M. Sternson & Karel Svoboda. Nature. 2009 Jan 18. 10.1038/nature07709 PubMed 19151697