Screen It™ CRISPR Cas9 Cleavage Detection Kit
Cat. No. | G990 | ||||||||||||||||||||||||||||
Name | Screen It™ CRISPR Cas9 Cleavage Detection Kit | ||||||||||||||||||||||||||||
Unit | 100 Reactions | ||||||||||||||||||||||||||||
Category | Cas Proteins & CRISPR Screening | ||||||||||||||||||||||||||||
Description |
abm’s Screen It™ CRISPR Cas9 Cleavage Detection Kit is a robust and precise RNA-dependent assay designed to detect successful insertions and deletions (indels) in genomic DNA following CRISPR-Cas9 editing. The kit can determine whether a clone is monoallelic (mutations in one copy), biallelic (mutations in both copies), or wild-type (unchanged), offering a key screening advantage over the standard T7E1 Surveyor Assay. The assay begins by amplifying the sgRNA target site from CRISPR-edited cells using PCR. The resulting amplicon is then cleaved by a ribonucleoprotein (RNP) complex, generating a distinct banding pattern that is easily resolved on an agarose gel. This clear visual output allows users to quickly and confidently assess editing outcomes. By streamlining the post-editing screening process, the Screen It™ Kit helps researchers save time and effort, providing reliable genotyping results for efficient identification of successfully edited clones.
Additional Materials Required
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Storage Condition | Store all components at -20°C. | ||||||||||||||||||||||||||||
Material Citation | If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. G990 |
What is the Screen It™ CRISPR Cas9 Cleavage Detection Kit used for? | |
This kit is designed to identify successful insertions or deletions (indels) in genomic DNA following CRISPR/Cas9 editing. It can determine if gene editing is monoallelic, biallelic, or unedited, providing valuable insight into the efficiency of the CRISPR process and the editing pattern.
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How does the Screen It™ kit work? | |
The Screen It™ CRISPR Cas9 Cleavage Detection Kit follows a streamlined process to identify successful CRISPR-induced insertions or deletions:
This kit helps efficiently determine the editing status (monoallelic, biallelic, or wild-type) of your CRISPR-edited samples.
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How do I prepare the PCR primers for this kit? | |
Primers should amplify the sgRNA target region and the Cas9 cut site, ensuring the cut site is not in the middle of the amplicon for clear cleavage patterns. If needed, abm offers a primer design service (Cat. No. C336) to help ensure that primers are optimized for your specific target and CRISPR experiment.
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How do I analyze the cleavage patterns on a gel? | |
To analyze the cleavage patterns on a gel:
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What do I need to synthesize sgRNA for use with this kit? | |
The kit includes a scaffold template and primer mix to transcribe sgRNA. Users need to input their sgRNA sequence into the kit for synthesis, which will be used to generate sgRNA suitable for the RNP cleavage reactions in the assay.
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Can I use this kit for high-throughput applications? | |
Yes, the kit’s streamlined protocol is designed for ease of use and rapid processing, making it suitable for high-throughput applications in genomic screening. It can help researchers quickly identify edited clones and save valuable time in CRISPR experiment workflows.
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What should I do if I see non-specific bands on the gel? | |
If non-specific bands appear, try using the touchdown PCR method to improve specificity. Adjusting PCR conditions can reduce non-specific amplification, helping ensure clear and accurate results for cleavage analysis.
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What is the source of this enzyme? | |
The enzyme is produced recombinantly in E. coli, which has been engineered to express the enzyme gene. While the original gene may come from another organism, all production and purification occur using E. coli under controlled conditions.
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