CRISPR02
CRISPR Nickase EMX1 Positive Control
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| Pack Size | SKU | Availability | Price |
|---|---|---|---|
| 1 set | Check Cart for Availability | $281.00 |
About This Item
NACRES:
NA.51
UNSPSC Code:
41106609
Concentration:
20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)
form
liquid
Quality Segment
packaging
pkg of 3 vials (50μL aliquot for each of the 3 kit components)
concentration
20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)
application(s)
CRISPR
shipped in
dry ice
storage temp.
−20°C
General description
Validated CRISPR site, which serves as an experimental control for the Cas9-D10A Nickase system. A three component positive control system consisting of a CMV-driven Cas9-D10A Nickase plasmid, a U6-driven guide RNA plasmid targeting the sense strand of the human EMX1 gene and a U6-driven guide RNA plasmid targeting the antisense strand of the human EMX1 gene.
Application
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
Biochem/physiol Actions
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.
Features and Benefits
Serves as an experimental control for the CRISPR editing workflow using Cas9-D10A Nickase. Allows for validation of your system with the CRSIPR/Cas9 system. A positive result in a miss-match detection assay will indicate validation of your system.
Physical form
Sigma U6-gRNA plasmid expressing a guide sequence to human EMX1 supplied at a concentration of 20ng/ul in 50ul. Sigma Cas9-D10A Nickase plasmid at a concentration of 20ng/ul in 50ul.
Preparation Note
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.
Other Notes
1 vial containing 1ug of a plasmid expressing the forward strand guide sequence. 1 vial containing 1ug of a plasmid expressing the reverse strand guide sequence. 1 vial containing 1ug of a plasmid expressing Cas9-D10A.
Typical transfection concentrations used in literature are in the ranges of >= 1.0ug/UL and <= 5uL of Cas9-D10A Nickase plasmids combined with >= 1.0ug/UL and <= 5uL of U6-gRNA plasmids. (All dosages above assume 0.5 to 1 million cells nucleofected)
Legal Information
1 of 1
This Item | |||
|---|---|---|---|
| concentration 20 ng/μL in TE buffer; DNA (1μg of plasmid DNA) | concentration 20 ng/μL in TE buffer; DNA (1μg of plasmid DNA) | concentration 20 ng/μL in TE buffer; DNA (1μg of plasmid DNA) | concentration 1x106 VP/ml (via p24 assay) |
| storage temp. −20°C | storage temp. −20°C | storage temp. −20°C | storage temp. −70°C |
| shipped in dry ice | shipped in dry ice | shipped in dry ice | shipped in dry ice |
| Quality Level 200 | Quality Level 200 | Quality Level 200 | Quality Level 200 |
| form liquid | form liquid | form liquid | form liquid |
| application(s) CRISPR | application(s) CRISPR | application(s) CRISPR | application(s) CRISPR |
Storage Class
12 - Non Combustible Liquids
wgk
WGK 1
flash_point_f
Not applicable
flash_point_c
Not applicable
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Global Trade Item Number
| SKU | GTIN |
|---|---|
| CRISPR02-1SET | 04061833533819 |