FNCAS9PROT
FnCas9 Protein
from Francisella novicida, recombinant, expressed in E. coli, 1X NLS
Synonym(s):
Francisella Novicida Cas9
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About This Item
UNSPSC Code:
12352202
NACRES:
NA.51
Recommended Products
recombinant
expressed in E. coli
Quality Level
Assay
≥95% (SDS-PAGE)
form
lyophilized powder
packaging
pkg of 1 kit (4 components)
application(s)
CRISPR
shipped in
wet ice
storage temp.
−20°C
Related Categories
General description
The purified FnCas9 protein (~190 kDa) is derived from the type II CRISPR/Cas system from the bacterium Francisella novicida and has been engineered to function using three molecular components: the FnCas9 protein, a crRNA of user-defined sequence, and a tracrRNA of constant sequence. The FnCas9 endonuclease can be programmed by changing the crRNA sequence to create a site-specific double-strand break (DSB) in plasmids or purified genomic DNA.
Application
Functional Genomics/Target Validation/Genome Editing/Molecular Biology/Molecular Cloning/Synthetic Biology
Features and Benefits
- Suitable for plasmid or large genomic DNA fragment cloning
- Results in 4-bp 5′ overhangs.
- Cuts purified plasmid DNA more efficiently than SpCas9
Packaging
pkg of 50 μg (≥ 300 pmol)
pkg of 250 μg (≥ 1500 pmol)
pkg of 250 μg (≥ 1500 pmol)
Components
Each kit consists of:
- one vial of FnCas9 recombinant protein
- one vial containing 1 mL of 1× Dilution buffer
- one vial containing 1 mL of Nuclease free water with glycerol
- one vial containing 1 mL of 10× Digestion Buffer
Principle
Conventional restriction enzymes such as EcoRI and HindIII can only cut specific short (6 bp) DNA sequences and are therefore limited to cloning small DNA fragments (typically ~4 kb). CRISPR-based endonculeases can cut user defined sequences of a longer length (~20 bp) making them more flexible and applicable to larger DNA fragments such as multigene bacterial operons or mammalian genomic DNA with native promoter, exon, and intron components.
Reconstitution
Lyophilized FnCas9 protein should be resuspended in the Reconstitution solution provided to desired concentration. Gently tap tube to completely dissolve lyophilized powder, incubate for 10 minutes on ice, and spin tube to bring material to bottom of tube.
Other Notes
Click here to order gRNA
Enter the 20bp genomic DNA target sequence (5′ to 3′) plus the adjacent 3′ PAM site. Under special instructions, indicate "FnCas9 crRNA"
Click here to order FnCas9 tracrRNA
Check out our other MISSION® Cas9 Proteins at SigmaAldrich.com/CRISPRproteins
Enter the 20bp genomic DNA target sequence (5′ to 3′) plus the adjacent 3′ PAM site. Under special instructions, indicate "FnCas9 crRNA"
Click here to order FnCas9 tracrRNA
Check out our other MISSION® Cas9 Proteins at SigmaAldrich.com/CRISPRproteins
Legal Information
MISSION is a registered trademark of Merck KGaA, Darmstadt, Germany
Kit Components Only
Product No.
Description
- FnCas9-NLS from Francisella novicida, expressed in Escherichia coli
- Dilution buffer for Cas9 proteins
- Reconstitution solution for Cas9 proteins
- Digestion buffer 10× for FnCas9
Signal Word
Warning
Hazard Statements
Precautionary Statements
Hazard Classifications
Eye Irrit. 2
Storage Class Code
11 - Combustible Solids
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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