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pAAV-ihSyn1-tTA
(Plasmid #99120)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 99120 Standard format: Plasmid sent in bacteria as agar stab 1 $65
AAV1 99120-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)
    NEB Stable
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    tTA
  • Insert Size (bp)
    2869
  • Mutation
    *(see below)
  • Promoter ihSyn1

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site KpnI (not destroyed)
  • 3′ cloning site EcoRI/HindIII (not destroyed)

Resource Information

Depositor Comments

*Note: The tTA in this plasmid was made from a reverse tetracycline transactivator (rtTA) by changing two amino acids (G19E and P56A). These two residue changes have been shown to be sufficient for the reverse phenotype (10.1073/pnas.130192197).

Information for AAV1 (Catalog # 99120-AAV1) ( Back to top )

Purpose

Ready-to-use AAV1 particles produced from pAAV-ihSyn1-tTA (#99120). In addition to the viral particles, you will also receive purified pAAV-ihSyn1-tTA plasmid DNA.

Expression of the tet-off transactivator from an inducible synapsin promoter. 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

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

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-ihSyn1-tTA was a gift from Viviana Gradinaru (Addgene plasmid # 99120 ; http://n2t.net/addgene:99120 ; RRID:Addgene_99120)

    For viral preps, please replace (Addgene plasmid # 99120) in the above sentence with: (Addgene viral prep # 99120-AAV1)

  • For your References section:

    Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems. Chan KY, Jang MJ, Yoo BB, Greenbaum A, Ravi N, Wu WL, Sanchez-Guardado L, Lois C, Mazmanian SK, Deverman BE, Gradinaru V. Nat Neurosci. 2017 Aug;20(8):1172-1179. doi: 10.1038/nn.4593. Epub 2017 Jun 26. 10.1038/nn.4593 PubMed 28671695