GeneArt™ Genomic Cleavage Detection Kit
100% knock-in using CRISPR-Cas9—try our new TrueTag Donor DNA Kits for fluorescent or epitope gene tagging.
GeneArt™ Genomic Cleavage Detection Kit
Invitrogen™

GeneArt™ Genomic Cleavage Detection Kit

The GeneArt Genomic Cleavage Detection Kit is a fast, T7 endonuclease I based method to quantify how well your genomeRead more
Have Questions?
Catalog number A24372
Price (USD)
259.00
Each
Add to cart
Price (USD)
259.00
Each
Add to cart
The GeneArt Genomic Cleavage Detection Kit is a fast, T7 endonuclease I based method to quantify how well your genome editing protocol causes insertions and deletions (indels) in the genome of your cell line. This is the quickest way to quantify and validate the best CRISPR-Cas9 gRNA, or the best TAL effector nuclease, for your cell engineering experiment. The GeneArt Genomic Cleavage Detection Kit provides a convenient, quick, and complete solution.

Using this T7 endonuclease I (T7EI) based method, you can quickly and confidently measure your on-target genome editing efficiency generated by non-homologous end joining (NHEJ) activity. The advantages of the GeneArt Genomic Cleavage Detection Kit are:

• Minimal hands-on time — PCR amplify from cell lysate, no DNA purification required
• Short protocol — four hours from cell harvest to quantified results
• Just add PCR primers — everything needed in one box. No separate PCR master mixes or DNA extraction kits to buy.
• Quantify editing efficiency directly from your gel — gel band density directly correlates to on-target indel formation

Obtain results the same day without waiting for sequencing results or performing complicated sequencing analysis.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Detection MethodPrimer-probe
FormLiquid
FormatKit
PolymeraseTaq Polymerase
Reaction SpeedFast
TechniqueCRISPR-Cas9, TAL Effector Nuclease
Product TypeGenomic Cleavage Detection Kit
Quantity20 reactions
Recognition SiteINDEL
Sufficient For20 Reactions
No. of Reactions20 Reactions
Unit SizeEach
Contents & Storage
Contains:

• 1 bottle Cell Lysis Buffer
• 1 tube Proteinase K
• 1 tube PCR Supermix
• 1 tube Water
• 1 tube T7E1 Detection Enzyme
• 1 tube T7E1 Detection Reaction Buffer
• 1 tube Control Template & Primers

Store all components at -5° to -30°C.

Frequently asked questions (FAQs)

What are TALs or TALENS?

TALs or TALENs are transcription activator-like effector nuclease proteins that are naturally occurring transcriptional activators secreted by Xanthomonas spp. into their plant hosts. GeneArt TALs are derived from Xathomonas TAL effectors, the DNA-binding domain of which consists of a variable number of amino acid repeats. Each repeat contains 33–35 amino acids and recognizes a single DNA base pair. The DNA recognition occurs via 2 hypervariable amino acid residues at positions 12 and 13 within each repeat, called repeat-variable di-residues (RVDs). TAL effector repeats can be assembled in modular fashion, varying the RVDs to create a TAL protein that recognizes a specific target DNA sequence.

What is CRISPR-STOP?

CRISPR-STOP is a method of inserting STOP codon sequences to generate knockouts.

Please refer to the following article: CRISPR-STOP: gene silencing through base-editing-induced nonsense mutations.

Find additional tips, troubleshooting help, and resources within our Genome Editing Support Center.

I am working with TALs and want to incorporate an effector domain that you do not carry. What should I do?

We do offer a multiple cloning site sequence in the place of the effector domain sequence for our TAL MCS entry vector. This option allows you to insert any protein-coding sequence, and allows your resulting TAL protein to deliver the effector in a sequence-specific manner anywhere in the genome. We also provide gene synthesis services to generate any effector domain for which you don't have a template.

I am trying to design my TAL but do not have a T at the 5´ end of the TAL effector. What should I do?

While our Invitrogen GeneArt Precision TALs required a T at the 5´end and 13-18 bp spacing between the forward and reverse TAL effectors for proper pairing of Fok1 nucleases, the Invitrogen GeneArt PerfectMatch TALs allow for targeting of any sequence across the genome and eliminates the 5´ T constraints. Additionally, the spacing between the two effectors is optimal at 15-16 bp.

The binding domain for TALs can be either 19 or 25 bp in length. Does one work better than the other?

The 19 bp binding domains perform better for the nucleases. The binding sites do not need to be the same size; however, best performance for the nucleases is with the 19 bp binding domains.

View all GeneArt™ Genomic Cleavage Detection Kit FAQs