추천 제품
포장
pkg of 8x25 μL (vials)
Quality Level
농도
5x108 VP/ml (via p24 assay)
응용 분야
CRISPR
배송 상태
dry ice
저장 온도
−70°C
일반 설명
CRISPR (clustered regularly interspaced short palindromic repeat) are repeated sequence of DNA. These are transcribed in a single transcript and later divided into short repeats.
애플리케이션
Functional Genomics/Screening /Target Validation
생화학적/생리학적 작용
Utilize the power of CRISPR Whole Genome Lentiviral Pools to knockout and screen every gene in the human genome. The Sigma Whole Human Genome Lentiviral CRISPR Pool is provided as 200 ul in 8 x 25 ul aliquots at a minimum titer of 5x108 viral particles/ml via p24.
특징 및 장점
- Use CRISPR nucleases to knockout protein-coding genes to assess their function
- Efficiently screen the whole human genome (16,000+ genes) at the bench-top without robotics or specialized equipment
- Increased flexibility consisting of 8 subpools at 23,000 clones per pool (184,000 clones total), from the previous 2 subpools (Gecko v2)
- Expanded gRNA coverage at 10 gRNAs per gene, from the previous 6 gRNAs per gene (Gecko v2)
- Numerous built-in enrichment and depletion controls allows researchers to confidently gauge the success of their pooled screening experiments
- 2 Vector System (pools are gRNA-only, Cas9 sold separately)
- Ease of optimization: Utilizes same vector system as Gecko v2 allowing for optimization one for both systems
- Minimize off-targeting: stringent gRNA design and tiling rules
제조 메모
Puro Kill Curve and Determining CFU (Colony Formation Unit) per mL.
Prior to performing a library-scale screening, two preliminary experiments must be conducted: (1) determine the sensitivity of target cell type to puromycin (kill curve), and (2) determine the functional titer of the lentivirus in your cell type by completing a colony-forming assay (measured in CFU/ml). The calculation of MOI (multiplicity of infection) should be based on the value of CFU. Different cell types vary in transduction efficiency and different lentiviral constructs do not behave identically, so it is critical to optimize your experimental conditions with control lentiviral CRISPR clones (available from Sigma) prior to performing your pooled experiment.
Prior to performing a library-scale screening, two preliminary experiments must be conducted: (1) determine the sensitivity of target cell type to puromycin (kill curve), and (2) determine the functional titer of the lentivirus in your cell type by completing a colony-forming assay (measured in CFU/ml). The calculation of MOI (multiplicity of infection) should be based on the value of CFU. Different cell types vary in transduction efficiency and different lentiviral constructs do not behave identically, so it is critical to optimize your experimental conditions with control lentiviral CRISPR clones (available from Sigma) prior to performing your pooled experiment.
기타 정보
This product is for R&D use only, not for drug, household, or other uses. Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices. Though the lentiviral transduction particles produced are replication incompetent, it is recommended that they be treated as Risk Group Level 2 (RGL-2) organisms in laboratory handling. Follow all published RGL-2 guidelines for laboratory handling and waste decontamination.
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 3
Flash Point (°F)
Not applicable
Flash Point (°C)
Not applicable
가장 최신 버전 중 하나를 선택하세요:
Campylobacter Ecology and Evolution (2014)
문서
Lentiviral vector systems prioritize safety features, with design precautions preventing replication. Good handling practices are essential for use.
Get tips for handling lentiviruses, optimizing experiment setup, titering lentivirus particles, and selecting helpful products for transduction.
프로토콜
Lentivirus versions of genome modification technologies support successful CRISPR, RNAi, and ORF experiments.
FACS sorts cells based on light scattering and fluorescence for objective cell analysis.
자사의 과학자팀은 생명 과학, 재료 과학, 화학 합성, 크로마토그래피, 분석 및 기타 많은 영역을 포함한 모든 과학 분야에 경험이 있습니다..
고객지원팀으로 연락바랍니다.