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px330-mcherry
(Plasmid #98750)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 98750 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pX330
  • Backbone manufacturer
    Zhang lab (Addgene plasmid #42230)
  • Modifications to backbone
    CMV- mcherry-pA amplified from mCherry-C1 (clontech) and inserted in to pX330 in NotI site
  • Vector type
    Mammalian Expression, CRISPR
  • Promoter U6
  • Selectable markers
    mCherry

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    NEB Stable
  • Copy number
    High Copy

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ sequencing primer CATCGTGGAACAGTACGAA
  • 3′ sequencing primer CGTTCACGGAGCCCTCCA
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

Because there exists a BbsI cleavage site on CMV-mcherry-pA cassette as well, it is suggested that the BbsI digested product need to be recovered directly instead of agarose gel purification.

User Instruction:
Digest the px330-mcherry plasmid with BbsⅠ enzyme. After that, run a small amount of the digest on an agarose gel electrophoresis to check the digestion efficiency. If the px330-mcherry plasmid has been well digested, recover the remaining digestion products using a DNA purification kit (such as Tiangen, DP214-03) for restriction enzyme cleanup. In our opinion, any commonly used DNA or PCR purification kit procedure would work well, but not agarose gel purification. We do not suggest combined digestion and ligation. For all other steps (including ligation of annealed oligos into this plasmid), refer to the Zhang lab protocol for pX330-U6-Chimeric_BB-CBh-hSpCas9 (Addgene plasmid #42230).

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:

    px330-mcherry was a gift from Jinsong Li (Addgene plasmid # 98750 ; http://n2t.net/addgene:98750 ; RRID:Addgene_98750)
  • For your References section:

    Correction of a genetic disease in mouse via use of CRISPR-Cas9. Wu Y, Liang D, Wang Y, Bai M, Tang W, Bao S, Yan Z, Li D, Li J. Cell Stem Cell. 2013 Dec 5;13(6):659-62. doi: 10.1016/j.stem.2013.10.016. 10.1016/j.stem.2013.10.016 PubMed 24315440