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Key Documents

FNCAS9PROT

Sigma-Aldrich

FnCas9 Protein

from Francisella novicida, recombinant, expressed in E. coli, 1X NLS

Synonyme(s) :

Francisella Novicida Cas9

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About This Item

Code UNSPSC :
12352202
Nomenclature NACRES :
NA.51

Produit recombinant

expressed in E. coli

Niveau de qualité

Pureté

≥95% (SDS-PAGE)

Forme

lyophilized powder

Conditionnement

pkg of 1 kit (4 components)

Application(s)

CRISPR

Conditions d'expédition

wet ice

Température de stockage

−20°C

Description générale

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

Caractéristiques et avantages

  • Suitable for plasmid or large genomic DNA fragment cloning
  • Results in 4-bp 5′ overhangs.
  • Cuts purified plasmid DNA more efficiently than SpCas9

Conditionnement

pkg of 50 μg (≥ 300 pmol)
pkg of 250 μg (≥ 1500 pmol)

Composants

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

Principe

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.

Autres remarques

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

Informations légales

MISSION is a registered trademark of Merck KGaA, Darmstadt, Germany

Composants de kit seuls

Réf. du produit
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

Pictogrammes

Exclamation mark

Mention d'avertissement

Warning

Mentions de danger

Classification des risques

Eye Irrit. 2

Code de la classe de stockage

11 - Combustible Solids


Certificats d'analyse (COA)

Recherchez un Certificats d'analyse (COA) en saisissant le numéro de lot du produit. Les numéros de lot figurent sur l'étiquette du produit après les mots "Lot" ou "Batch".

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Protocoles

Combine guaranteed sgRNAs with our comprehensive range of CRISPR products and tools, including Cas9 and delivery reagents, for efficient genome modification with higher specificity.

Contenu apparenté

Combine guaranteed sgRNAs with our comprehensive range of CRISPR products and tools, including Cas9 and delivery reagents, for efficient genome modification with higher specificity.

Watch Cas9-GFP proteins work inside your cells and deliver the powerful nuclease editing you have come to trust and expect. MISSION™ SpCas9-GFP and eSpCas9-GFP proteins are multi-functional enzymes with high on-target cleavage efficiency fused to Enhanced Green Fluorescent Protein (EGFP) for real-time observation of Cas9 delivery and clearance from cells.

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