B-HiFi SpCas9
(Plasmid
#207362)
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PurposeExpression plasmid for human codon-optimized increased-fidelity (i.e. high-fidelity) SpCas9 variant
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Depositing Lab
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Depositing OrganizationHUN-REN Research Centre for Natural Sciences
Located in Hungary -
Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 207362 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 207362-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 207362-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepX330-like (without U6-sgRNA coding sequence)
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Vector typeMammalian Expression, CRISPR
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 nameB-HiFi SpCas9
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SpeciesS. pyogenes
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Insert Size (bp)4251
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MutationR691A, amino acids 1005-1013 replaced with two glycine
- Promoter CBh
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Tag
/ Fusion Protein
- 3xFLAG (N terminal on insert)
Cloning Information
- Cloning method Gibson Cloning
Resource Information
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
Please visit https://doi.org/10.1101/2022.08.16.504097 for bioRxiv preprint
DNA (Catalog # 207362-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
-
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 # 207362-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:
B-HiFi SpCas9 was a gift from Ervin Welker (Addgene plasmid # 207362 ; http://n2t.net/addgene:207362 ; RRID:Addgene_207362) -
For your References section:
A cleavage rule for selection of increased-fidelity SpCas9 variants with high efficiency and no detectable off-targets. Kulcsar PI, Talas A, Ligeti Z, Toth E, Rakvacs Z, Bartos Z, Krausz SL, Welker A, Vegi VL, Huszar K, Welker E. Nat Commun. 2023 Sep 16;14(1):5746. doi: 10.1038/s41467-023-41393-5. 10.1038/s41467-023-41393-5 PubMed 37717069
