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PurposeCRISPR/Cas9 system gRNA cloning
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Depositing Lab
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Depositing OrganizationFudan University
Located in China -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 61089 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 61089-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 61089-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepUC19
- Backbone size w/o insert (bp) 2636
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Vector typeMammalian Expression, CRISPR ; /Cas9 gRNA cloning vector
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameH1 promoter; gRNA sequences
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SpeciesH. sapiens (human)
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Insert Size (bp)346
- Promoter H1
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site EcoRI (not destroyed)
- 3′ cloning site HindIII (not destroyed)
- 5′ sequencing primer M13-F20
- 3′ sequencing primer M13R
- (Common Sequencing Primers)
Resource Information
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Addgene Notes
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A portion of this plasmid was derived from a plasmid made byNS
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
As Ampicillin gene and H1 promoter contain BsaI sites, we mutated the sequence of Amp gene (G1601C, not change amino acid), and mutated H1 promoter from GAGACC to GAGGACC to eliminate the BsaI internal site.
DNA (Catalog # 61089-D50)
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
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Academic/Nonprofit Terms
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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.
DNA (Catalog # 61089-D100)
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.
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.
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For your Materials & Methods section:
pUC-H1-gRNA was a gift from Shenglin Huang (Addgene plasmid # 61089 ; http://n2t.net/addgene:61089 ; RRID:Addgene_61089) -
For your References section:
Precise gene deletion and replacement using the CRISPR/Cas9 system in human cells. Zheng Q, Cai X, Tan MH, Schaffert S, Arnold CP, Gong X, Chen CZ, Huang S. Biotechniques. 2014 Sep 1;57(3):115-24. doi: 10.2144/000114196. eCollection 2014. 10.2144/000114196 PubMed 25209046