추천 제품
재조합
expressed in E. coli
Quality Level
분석
≥95% (SDS-PAGE)
양식
lyophilized powder
포장
pkg of 1 kit (4 components)
응용 분야
CRISPR
배송 상태
wet ice
저장 온도
−20°C
일반 설명
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.
애플리케이션
Functional Genomics/Target Validation/Genome Editing/Molecular Biology/Molecular Cloning/Synthetic Biology
특징 및 장점
- Suitable for plasmid or large genomic DNA fragment cloning
- Results in 4-bp 5′ overhangs.
- Cuts purified plasmid DNA more efficiently than SpCas9
포장
pkg of 50 μg (≥ 300 pmol)
pkg of 250 μg (≥ 1500 pmol)
pkg of 250 μg (≥ 1500 pmol)
성분
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
원리
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.
재구성
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.
기타 정보
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
법적 정보
MISSION is a registered trademark of Merck KGaA, Darmstadt, Germany
키트 구성품 전용
제품 번호
설명
- FnCas9-NLS from Francisella novicida, expressed in Escherichia coli
- Dilution buffer for Cas9 proteins
- Reconstitution solution for Cas9 proteins
- Digestion buffer 10× for FnCas9
신호어
Warning
유해 및 위험 성명서
Hazard Classifications
Eye Irrit. 2
Storage Class Code
11 - Combustible Solids
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프로토콜
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