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HWGCRISPR

Sigma-Aldrich

Sigma Whole Human Genome Lentiviral CRISPR Pool

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About This Item

Código UNSPSC:
12352200
NACRES:
NA.51

embalagem

pkg of 8x25 μL (vials)

Nível de qualidade

200

concentração

5x108  VP/ml (via p24 assay)

aplicação(ões)

CRISPR

Condições de expedição

dry ice

temperatura de armazenamento

−70°C

Categorias relacionadas

Descrição geral

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.

Aplicação

Functional Genomics/Screening /Target Validation

Ações bioquímicas/fisiológicas

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.

Características e benefícios

  • 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

Nota de preparo

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.

Outras notas

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.

Informações legais

CRISPR Use License Agreement

Lentiviral and WPRE License Agreements

Código de classe de armazenamento

12 - Non Combustible Liquids

Classe de risco de água (WGK)

WGK 3

Ponto de fulgor (°F)

Not applicable

Ponto de fulgor (°C)

Not applicable

Certificados de análise (COA)

Busque Certificados de análise (COA) digitando o Número do Lote do produto. Os números de lote e remessa podem ser encontrados no rótulo de um produto após a palavra “Lot” ou “Batch”.

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Campylobacter Ecology and Evolution (2014)

Artigos

Safe Lentivirus Handling

Our lentiviral vector systems are developed with enhanced safety features. Numerous precautions are in place in the design of our lentiviruses to prevent replication. Good handling practices are a must.

Successful Transduction Using Lentivirus

Successful targeting relies on optimizing key sensitive steps in the process, including lentiviral transduction. Below are some helpful handling and titration tips from our R&D lentiviral experts.

Protocolos

CRISPR Lentiviral Screening: Antibiotic Selection

You are not alone designing successful CRISPR, RNAi, and ORF experiments. Sigma-Aldrich was the first company to commercially offer lentivirus versions of targeted genome modification technologies and has the expertise and commitment to support new generations of scientists.

FACS Titration of Lentivirus Protocol

FACS (Fluorescence-Activated Cell Sorting) provides a method for sorting a mixed population of cells into two or more groups, one cell at a time, based on the specific light scattering and fluorescence of each cell. This method provides fast, objective, and quantitative recording of fluorescent signals from individual cells.

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