Pluripotent Stem Cell (PSC) Gene Editing Demo Kit

Catalog No.	Quantity
A43203	1 Kit

Catalog No. A43203 Supplier Gibco Supplier No. A43203

Description

The Pluripotent Stem Cell (PSC) Gene Editing Demo Kit offers the critical reagents for successfully generating and detecting knock-outs and point mutations in your cell line of interest.

• Reduce effort and cycle times
• Generate pools with high editing efficiency (up to 80% KO and 40% HDR)
• Quantitate editing efficiency
• Isolate clonal lines easily

Includes:

• StemFlex Medium (500 mL), comprised of:
  • StemFlex Basal Medium (450 mL)
  • StemFlex Supplement (50 mL)
• rhLaminin-521 (0.1 mg)
• RevitaCell Supplement (5 mL)
• TrueCut Cas9 Protein v2 (10 μg)
• TrueGuide sgRNA Positive Control, HPRT1
• GeneArt Genomic Cleavage Detection Kit
• Manual/protocol (electronic)
• Neon demo upon request

Efficient genome editing

TrueCut Cas9 protein v2 features include:

• High DNA cleavage and homology driven repair
• Complex with gRNAs into RNPs
• Deliver RNPs with Neon system or Lipofectamine Stem reagent

Superior single-cell survival

• The StemFlex/Laminin-521 system with RevitaCell supplement enhances single-cell survival afterelectroporation and single cell sorting. Improved cell survival after electroporation reduces timelines, and the ability to single-cell sort hiPSCs reduces clonal isolation effort and timelines dramatically.

Rapid detection

• The GeneArt Genomic Cleavage Detection Kit is a fast, PCR-based method to measure how well your genome editing tool forms insertion and deletion (indel) mutations. Directly PCR amplify your pool of edited cells with primers specific to your targeted region, run the cleavage assay, and determine efficiency, all in an afternoon.

Specifications

• Cell Type: Embryonic Stem Cells (ESCs), Induced Pluripotent Stem Cells (iPSCs) 
• Product Type: Pluripotent Stem Cell (PSC) Gene Editing Demo Kit
• Quantity: 1 Kit
• Content And Storage: • StemFlex Medium (500 mL), comprised of:
     --StemFlex Basal Medium (450 mL, 2–8°C)
     --StemFlex Supplement (50 mL, -5 to -30°C)
  • rhLaminin-521 (0.1 mg, -5 to -30°C)
  • RevitaCell Supplement (5 mL, -5 to -30°C)
  • TrueCut Cas9 Protein v2 (10 μg, -5 to -30°C)
  • TrueGuide Positive Control Guide sgRNA, HPRT1 (3 nmol; store dried RNA at room temperature, store resuspended RNA at -5 to -30°C; resuspend in nuclease-free water or TE)
  • Genomic DNA Cleavage Detection Kit (1 kit, -5 to -30°C)
  
  

Frequently Asked Questions (FAQs)

The Pluripotent Stem Cell (PSC) Gene Editing Demo Kit contains Cas9 protein and gRNA as editing tools. Can I use my all-in-one vectors instead?

The kit is optimized to work with Cas9 protein and gRNA (either synthetic or IVT gRNA). This combination provides the highest editing efficiency in hIPSC. After electroporation optimization, DNA-based methods may still work at lower efficiencies however, and the matrix and medium in the kit can certainly be used to maintain cells after delivery of DNA.

This Pluripotent Stem Cell (PSC) Gene Editing Demo Kit includes StemFlex Medium, however I have all my cells growing in Essential 8 Medium. Can I use Essential 8 Medium instead?

Survival after electroporation is much poorer when using Essential 8 Medium compared to that observed with StemFlex Medium. If Essential 8 Medium is important for downstream processes, cells can be adapted to StemFlex Medium for the genome editing portion and then adapted back to Essential 8 Medium for the remainder of the work.

With the Pluripotent Stem Cell (PSC) Gene Editing Demo Kit, why do you recommend using electroporation? What about lipofection with, for example, Lipofectamine Stem Transfection Reagent?

Electroporation provides superior delivery of genome editing tools, resulting in higher editing efficiencies.

For Research Use Only. Not for use in diagnostic procedures.