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CRISPR-Q Sanger Primers (10 µM)
Cat. No. / ID: 232104
Two vials containing lyophilized Sanger sequencing primers for analyzing PCR products generated with the corresponding CRISPR-Q Custom PCR Assay; 10 µM
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CRISPR-Q Sanger Primers are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
Configure at GeneGlobe
Find or custom design the right target-specific assays and panels to research your biological targets of interest.
Features
- Highly specific primers for successful sequencing and analysis
- Simple-to-use primer design tool requires no design expertise
- Sanger analysis tool provides fast and easy analysis of sequencing traces and calculation of editing efficiency
Product Details
CRISPR-Q Sanger Primers enable rapid characterization of CRISPR-based genome editing events involving human, mouse or rat cells. These primers are designed for use with the QIAprep& CRISPR Kit with an innovative workflow that combines liquid-based sample preparation with Sanger sequencing-based analysis 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
CRISPR-Q Sanger Primers and the corresponding CRISPR-Q Sanger Sequencing Analysis Tool allow you to calculate and visualize the efficiency of your editing event with speed, convenience and accuracy (see figure The CRISPR-Q Sanger Primers successfully sequence target regions of interest).
See figures
Principle
The easy-to-use CRISPR assay design tool, which is available in GeneGlobe, requires no primer design expertise and provides full support for human, mouse and rat targets. Just enter your chromosome number, cut site and guide RNA sequence, and our advanced design algorithm will do the rest. The design is based on the genomic location of the cut and generates three robust PCR primer sets and corresponding CRISPR-Q Sanger Primers that enable you to analyze the specified region with high success. The CRISPR-Q Sanger analysis tool, also available in GeneGlobe, enables calculation and visualization of your CRISPR editing events.
Procedure
The QIAprep& 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 (see figure The QIAprep& CRISPR Kit workflow). First, the target region is amplified using the QIAprep& CRISPR Kit and CRISPR-Q Custom PCR Assays. Following purification of the PCR product, it is sequenced using the CRISPR-Q Sanger Primers and the sequence traces are analyzed using the CRISPR-Q Sanger Sequencing Analysis tool, which is available in GeneGlobe.
Primers for Sanger sequencing of genomic regions of interest
CRISPR-Q Sanger Primers can be easily designed and ordered using the intuitive custom builder tool, which is also available in GeneGlobe. This tool generates several target-specific primers based on the genomic location and the sequence of the guide RNA used for your particular CRISPR gene editing events.
See figures
Applications
The QIAprep& 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)
Supporting data and figures
The QIAprep& CRISPR Kit workflow.
The new CRISPR workflow combines fast, liquid-based sample preparation with PCR and Sanger sequencing-based analysis for checking the gene editing efficiency.
Resources
Certificates of Analysis (1)
FAQ
What is the minimum cell number needed for cell lysis?
Is the QIAprep& CRISPR Kit also applicable to other gene-editing technologies such as TALENs and ZFN?
Which database versions are used for CRISPR-Q Custom PCR Assay and CRISPR-Q Sanger Primers design?
For the Sanger analysis tool, the customer needs to upload two .abi1 files of forward and reverse sequences? Is there any additional info that needs to be provided?
Are the PCR products generated with CRISPR-Q Custom PCR Assays only applicable to analysis of editing efficiency by Sanger sequencing?
Is there something to consider when working with transduced cells treated with Polybrene?
Are the CRISPR-Q PCR Assays and the CRISPR-Q Sanger Primers on GeneGlobe validated?
For sequencing application, does the lysate require purification?
Can the new CRISPR PCR Assays be used for dPCR on the QIAcuity? Do you have data or a protocol?
Is there anything to consider when processing cultured cells on coated cultivation dishes? Are additional purification steps required to eliminate or reduce coating reagents such as Poly-L-Lysine?