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重组
expressed in E. coli
质量水平
方案
≥95% (SDS-PAGE)
表单
lyophilized powder
包装
pkg of 1 kit (4 components)
应用
CRISPR
运输
wet ice
储存温度
−20°C
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)
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
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
仅试剂盒组分
产品编号
说明
- FnCas9-NLS from Francisella novicida, expressed in Escherichia coli
- Dilution buffer for Cas9 proteins
- Reconstitution solution for Cas9 proteins
- Digestion buffer 10× for FnCas9
警示用语:
Warning
危险声明
危险分类
Eye Irrit. 2
储存分类代码
11 - Combustible Solids
法规信息
常规特殊物品
历史批次信息供参考:
分析证书(COA)
实验方案
Combine guaranteed sgRNAs with our comprehensive range of CRISPR products and tools, including Cas9 and delivery reagents, for efficient genome modification with higher specificity.
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