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pCAG-T3-eSpCAS9-NG-pA
(Plasmid #131466)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 131466 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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DNA 131466-D50 50 μg of DNA in Tris buffer $495
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DNA 131466-D100 100 μg of DNA in Tris buffer $585
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Backbone

  • Vector backbone
    pCAGGS
  • Total vector size (bp) 9293
  • Vector type
    Mammalian Expression, CRISPR

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    Codon optimized eSpCas9-NG
  • Species
    Synthetic
  • Insert Size (bp)
    4218
  • Tags / Fusion Proteins
    • Flag (N terminal on insert)
    • NLS (N terminal on insert)
    • NLS (C terminal on insert)
    • polyA (C terminal on insert)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry

Trademarks:

  • Zeocin® is an InvivoGen trademark.

Depositor Comments

This plasmid vector was constructed by mutation PCR of pCAG-T3-hCAS9-pA plasmid vector (Addgene plasmid 48625) according to the amino acid sequences of SpCas9-NG shown in previous study (Nishimasu H et al., Engineered CRISPR-Cas9 nuclease with expanded targeting space. Science, 2018) with high-fidelity eSp variant shown in previous study (Slaymaker IM et al, Rationally engineered Cas9 nucleases with improved specificity. Science, 2016).

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

  • Amount 50 μg
  • Concentration 1 μg/μL
  • Pricing $495 USD
  • Storage 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

  • Amount 100 μg
  • Concentration 1 μg/μL
  • Pricing $585 USD
  • Storage 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

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:

    pCAG-T3-eSpCAS9-NG-pA was a gift from Wataru Fujii (Addgene plasmid # 131466 ; http://n2t.net/addgene:131466 ; RRID:Addgene_131466)

  • For your References section:

    Generation of genetically modified mice using SpCas9-NG engineered nuclease. Fujii W, Ito H, Kanke T, Ikeda A, Sugiura K, Naito K. Sci Rep. 2019 Sep 9;9(1):12878. doi: 10.1038/s41598-019-49394-5. 10.1038/s41598-019-49394-5 PubMed 31501500
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