CRISPR31
CRISPR Non-Target Negative Control Plasmid for Bacteria
About This Item
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form
liquid
packaging
vial of 50 μL
concentration
20 ng/μL in TE buffer; DNA (1μg of purified plasmid DNA)
technique(s)
microbiological culture: suitable
application(s)
CRISPR
genome editing
Promoter
Promoter activity: constitutive
shipped in
dry ice
storage temp.
−20°C
1 of 4
This Item | CRISPR30 | CAS9BAC1P | CRISPR19 |
---|---|---|---|
liquid | liquid | liquid | liquid |
microbiological culture: suitable | microbiological culture: suitable | microbiological culture: suitable | - |
dry ice | dry ice | dry ice | dry ice |
−20°C | −20°C | −20°C | −20°C |
vial of 50 μL | vial of 50 μL | vial of 50 μL | vial of 50 μL |
General description
Here we present a novel dual-vector CRISPR/Cas-mediated λ-Red system for improved recombineering in E. coli. Our system is shown to facilitate homology-directed repair of DSBs created by Cas9 endonuclease, enabling genetic alterations through chromosomal integration of a donor DNA.
This plasmid is to be used in combination with the Cas9 Lambda Red homologous recombination plasmid for E. coli (CAS9BAC1P) as the negative control for your custom gene editing experiment. The custom gRNA (CRISPRBACD) can be designed and ordered through https://www.sigmaaldrich.com/pc/ui/genomics-home/customcrispr
The CRISPR Non-Target Negative Control Plasmid for Bacteria (CRISPR31) contains a non-targeting spacer expressed constitutively from a J23119 promoter, a ampicillin resistance marker, a pBR322 origin of replication, and a sacB gene from Bacillus subtilis for counter-selection-based curing.
Application
- HR-mediated recombineering for mutation or SNP analysis
- Creation of HR-mediated knock-in cell lines with promoters, fusion tags, or reporters integrated into endogenous genes
- Creation of gene knockouts in E. coli cell lines
Strain Optimization
Features and Benefits
Markerless: does not require antibiotic resistance marker insertion
Scarless: no scar sequences from marker excision which often cause off-target recombination
Multiplexing: multiple custom gRNA sequences can be used at a time
Principle
Legal Information
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 2
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
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Articles
In this article, we present an application of our novel E. coli CRISPR/Cas-mediated Lambda-Red (λ-Red) homologous recombination (HR) vector system, which facilitates gene editing through the homology-directed repair (HDR) of double-stranded DNA breaks (DSBs) created by Cas9 endonuclease, using either ssDNA or dsDNA as an editing template.
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