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px458_Conc2
(Plasmid #134449)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 134449 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pSpCas9(BB)-2A-GFP (PX458)
  • Backbone manufacturer
    Feng Zhang
  • Modifications to backbone
    The Backbone vector was PciI/XbaI digested and an amplified region from 3 gRNA concatemer vector (#84880) was inserted, which encodes for two U6 promotors.
  • Vector type
    Mammalian Expression, CRISPR
  • Promoter U6
  • Selectable markers
    GFP

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    NEB Stable
  • Copy number
    Unknown

Cloning Information

  • Cloning method Restriction Enzyme

Resource Information

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

The sgRNAs can get introduced by golden gate cloning like described in Andersson-Rolf A et al.: Simultaneous paralogue knockout using a CRISPR-concatemer in mouse small intestinal organoids. Dev Biol. 2016. doi: 10.1016/j.ydbio.2016.10.016. For more information see protocol.

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:

    px458_Conc2 was a gift from Daniel Stange (Addgene plasmid # 134449 ; http://n2t.net/addgene:134449 ; RRID:Addgene_134449)
  • For your References section:

    Universal and Efficient Electroporation Protocol for Genetic Engineering of Gastrointestinal Organoids. Gaebler AM, Hennig A, Buczolich K, Weitz J, Welsch T, Stange DE, Pape K. J Vis Exp. 2020 Feb 18;(156). doi: 10.3791/60704. 10.3791/60704 PubMed 32150173