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HWGCRISPR

Sigma-Aldrich

Sigma Whole Human Genome Lentiviral CRISPR Pool

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About This Item

Code UNSPSC :
12352200
Nomenclature NACRES :
NA.51

Conditionnement

pkg of 8x25 μL (vials)

Niveau de qualité

200

Concentration

5x108  VP/ml (via p24 assay)

Application(s)

CRISPR

Conditions d'expédition

dry ice

Température de stockage

−70°C

Catégories apparentées

Description générale

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.

Application

Functional Genomics/Screening /Target Validation

Actions biochimiques/physiologiques

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.

Caractéristiques et avantages

  • 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

Notes préparatoires

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.

Autres remarques

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.

Informations légales

CRISPR Use License Agreement

Lentiviral and WPRE License Agreements

Code de la classe de stockage

12 - Non Combustible Liquids

Classe de danger pour l'eau (WGK)

WGK 3

Point d'éclair (°F)

Not applicable

Point d'éclair (°C)

Not applicable

Certificats d'analyse (COA)

Recherchez un Certificats d'analyse (COA) en saisissant le numéro de lot du produit. Les numéros de lot figurent sur l'étiquette du produit après les mots "Lot" ou "Batch".

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Campylobacter Ecology and Evolution (2014)

Articles

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.

Protocoles

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.

Notre équipe de scientifiques dispose d'une expérience dans tous les secteurs de la recherche, notamment en sciences de la vie, science des matériaux, synthèse chimique, chromatographie, analyse et dans de nombreux autres domaines..

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