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Merck

CRISPR08

Sigma-Aldrich

CRISPR UNIVERSAL NEGATIVE CONTROL 3

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About This Item

Código UNSPSC:
41106609
NACRES:
NA.51

formulario

liquid

Nivel de calidad

envase

vial of 50 μL

concentración

20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)

aplicaciones

CRISPR

Condiciones de envío

dry ice

temp. de almacenamiento

−20°C

Descripción general

Universal negative control CRISPRs have been designed not to recognize any sequence in the human, mouse or rat genome. A single vector format is provided which includes the Cas9 expression cassette and gRNA sequence. This vector includes GFP which is co-expressed from the same mRNA as the Cas9 protein via a 2A peptide linkage and enables for tracking of transfection efficiency or enrichment in cell populations via FACS.

Aplicación

Functional Genomics/Target Validation
  • Creation of gene knockouts in cell lines
  • Creation of knock-in cell lines with promoters, fusion tags or reporters integrated into endogenous genes

Características y beneficios

Allows for expression of Cas9 and GFP without specific targeting of the CRISPR/Cas9 system.

Componentes

1 vial containing 1ug of U6-gRNA/CMV-Cas9-GFP plasmid expressing a non-targeting guide sequence.

Principio

CRISPR/Cas systems are employed by bacteria and archaea as a defense against invading viruses and plasmids. Recently, the type II CRISPR/Cas system from the bacterium Streptococcus pyogenes has been engineered to function in eukaryotic systems using two molecular components: a single Cas9 protein and a non-coding guide RNA (gRNA). The Cas9 endonuclease can be programmed with a single gRNA, directing a DNA double-strand break (DSB) at a desired genomic location. Similar to DSBs induced by zinc finger nucleases (ZFNs), the cell then activates endogenous DNA repair processes, either non-homologous end joining (NHEJ) or homology-directed repair (HDR), to heal the targeted DSB.

Forma física

Sigma U6-gRNA/CMV-Cas9-GFP plasmid expressing a non-targeting guide sequence supplied at a concentration of 20ng/ul in 50ul.

Nota de preparación

Sigma CRISPR plasmid products are delivered as mini-prep aliquots, which may not be suitable for transfection into particular cell types. For best results, we advise maxi-prepping plasmids using endotoxin-free DNA purification kits prior to transfection.

Otras notas

Typical transfection concentrations used in literature are in the ranges of 2.0 to 8.0ug of the single vector expressing the guideRNA and Cas9 . (All dosages above assume 0.5 to 1 million cells nucleofected)

Código de clase de almacenamiento

12 - Non Combustible Liquids

Clase de riesgo para el agua (WGK)

WGK 1

Punto de inflamabilidad (°F)

Not applicable

Punto de inflamabilidad (°C)

Not applicable


Certificados de análisis (COA)

Busque Certificados de análisis (COA) introduciendo el número de lote del producto. Los números de lote se encuentran en la etiqueta del producto después de las palabras «Lot» o «Batch»

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Artículos

CRISPR endonucleases have shown wide variation in their activity, even among multiple CRISPRs designed within close genomic proximity.

View experimental data showing crispr/cas expression and enrichment using FACS

Protocolos

Learn about CRISPR Cas9, what it is and how it works. CRISPR is a new, affordable genome editing tool enabling access to genome editing for all.

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