FnCas9 Protein

半岛bd体育手机客户端 说明

一般描述

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.

应用

Functional Genomics/Target Validation/Genome Editing/Molecular Biology/Molecular Cloning/Synthetic Biology

特点和优势

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

包装

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

组分

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

原理

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.

重悬

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.

其他说明

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

法律信息

CRISPR Use License Agreement
MISSION is a registered trademark of sigma-aldrich Co. LLC

半岛bd体育手机客户端 性质

质量水平	200
重组	expressed inE. coli
测定	≥95% (SDS-PAGE)
形式	lyophilized powder
包装	pkg of 1 kit (4 components)
application(s)	CRISPR
运输	wet ice
储存温度	?20℃

组份列表

组份不可单独销售

货号	组份
	FnCas9-NLS fromFrancisella novicida,expressed inEscherichia coli
	Dilution buffer for Cas9 proteins
	Reconstitution solution for Cas9 proteins
	Digestion buffer 10× for FnCas9

安全信息

象形图
警示用语：	Warning
危险声明	H319
预防措施声明	P264 - P280 - P305 + P351 + P338 - P337 + P313
危险分类	Eye Irrit. 2
储存分类代码	11 - Combustible Solids
WGK	WGK 3