HWGCRISPR

Sigma Whole Human Genome Lentiviral CRISPR Pool

• UNSPSC kód: 41106609
• NACRES: NA.51

Tulajdonságok

• kiszerelés: pkg of 8x25 μL (vials)
• Minőségi szint: 200
• koncentráció: 5x10⁸ VP/ml (via p24 assay)
• alkalmazás(ok): CRISPR
• kiszállítva: dry ice
• tárolási hőmérséklet: −70°C

Leírás

Általános leírás

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.

Alkalmazás

Functional Genomics/Screening /Target Validation

Biokémiai/fiziológiai hatások

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.

Tulajdonságok és előnyök

• 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

Elkészítési megjegyzés

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.

Egyéb megjegyzések

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.

Jogi információk

CRISPR Use License Agreement

Lentiviral and WPRE License Agreements

Biztonsági adatok

• Tárolási osztály kódja: 12 - Non Combustible Liquids
• WGK: WGK 3
• Lobbanási pont (F): Not applicable
• Lobbanási pont (C): Not applicable