For PCR-based characterization and confirmation of CRISPR editing events in human, mouse, or rat cells

Features

Optimized for usage with the QIAprep&;amp CRISPR Kit

Fully customized for your region of interest

in silico validation ensures high success rate

Design available for human, mouse and rat genomes

PCR product obtained is compatible with T7 and Sanger sequencing analysis

Product Details

CRISPR-Q Custom PCR Assays enable rapid and sensitive characterization of CRISPR-based genome editing events involving human, mouse or rat cells. These assays are designed for use with the QIAprep&;amp CRISPR Kit with an innovative workflow that combines liquid-based sample preparation with end-point PCR detection of your region of interest. The CRISPR-Q Sanger Sequencing Analysis tool, which is available in GeneGlobe, enables quick and convenient analysis to complete the entire CRISPR editing detection workflow.

Performance

The CRISPR-Q Custom PCR Assays and the QIAprep&;amp CRISPR Kit provide sensitive detection from a wide cell input range . Our advanced assay design algorithm provides full support for human, mouse and rat targets and generates robust primer sets that amplify the specified targets with a high success rate.

Principle

CRISPR-Q Custom PCR Assays allow you to characterize CRISPR editing events in cultured human, mouse or rat cells. The easy-to-use assay design tool is available in GeneGlobe: just enter your chromosome number, cut site and guide RNA sequence, and our advanced design algorithm will do the rest. It checks for the presence of gRNA sequence in the PCR product to ensure amplification of the correct region of interest. It also checks for off-target amplification to ensure that only the specific target is amplified and to provide an optimal user experience.

Three primer sets flanking the endonuclease cut site are designed for each target. The resulting PCR amplicons are 400–600 bp long with at least 150 bp from the cut site. The amplicons are compatible with various downstream analysis methods such as Sanger sequencing or mismatch detection assays.

Procedure

The QIAprep&;amp CRISPR Kit offers a streamlined workflow, which combines a liquid-based sample preparation step that can be completed in only 25 minutes with sensitive PCR and Sanger sequencing detection . 

CRISPR-Q Custom PCR Assays can be directly used for target amplification form lysed cultured cells derived from human, mouse or rat without additional optimization of the PCR. The PCR primers are designed so that the resulting PCR product can be used for various downstream analyses.

Assays for amplifying genomic regions of interest

CRISPR-Q Custom PCR Assays can be easily designed and ordered for human, mouse or rat gene targets using the intuitive custom builder tool available in GeneGlobe at www.geneglobe.com/customize/crispr/ (see figure  CRISPR assay design is optimized for human, mouse and rat gene targets). The custom builder tool generates several target-specific assays based on the genomic location and the sequence of the guide RNA (gRNA) used for your particular CRISPR gene editing events.

Applications

The QIAprep&;amp CRISPR Kit and corresponding CRISPR-Q Custom PCR Assays and CRISPR-Q Sanger Primers are well-suited for practically any researcher who needs to characterize CRISPR-based editing events:

Characterization of gene editing events

Functional studies including gene knock-out or insertion

Base editing

Genome screening (CRISPRi libraries, reversible knockdowns)