High-Throughput Genotyping (CRISPR Validation)

 

High-throughput genotyping is a next generation sequencing (NGS) assay ideal for ultra-sensitive, high-throughput, sample-to-answer genotyping projects, enabling validation of targeted CRISPR gene editing experiments. Recent advancements in genome editing techniques, namely CRISPR/Cas9, have greatly facilitated cell line engineering, gene targeting, and the development of transgenic models. However, identification of correctly targeted cells remains a significant bottleneck for large-scale projects.

GENEWIZ genoTYPER-NEXT™ from Azenta Life Sciences is a proprietary tool more effective and efficient than T7E1, TIDE, and IDAA assays for screening targeted cell lines. We also offer targeted, whole genome, and amplicon sequencing NGS solutions to aid on- and off-target analysis of genome editing.

 

 

What is high-throughput genotyping?

High-throughput genotyping is the rapid analysis of genetic markers or gene variations in a sample, often with tens to hundreds of samples simultaneously. This method is a valuable tool for researchers conducting genome-wide association studies (GWAS), population genetics studies, pharmacogenomics, and research involving genetic risk factors and responses to drug therapies.

 

How do you validate CRISPR gene editing?

CRISPR gene editing is validated by polymerase chain reaction (PCR), Sanger sequencing, or NGS. These techniques are used to identify and verify gene modifications within the edited DNA sample.

 

What is the target validation of CRISPR-Cas9?

The target validation of CRISPR-Cas9 is the confirmation of selected guide RNA (gRNA) sequences and its specificity to the gene of interest. This method involves analyzing other genomic loci that share sequence similarity with the chosen gRNA and comparing it to the non-targeted controls (gRNA targeting a non-functional region) to ensure that the gRNA predominantly targets the intended genomic locus. The functional consequences of gene editing,such gene knockout and subsequent absence of protein expression, are confirmed and quantified via quantitative PCR (qPCR) or deep sequencing.

genoTYPER-NEXT: HIGH-THROUGHPUT ASSAY FOR CRISPR VALIDATION

1. SUBMIT SAMPLES

Submit CRISPR-edited cell lines in 96-well plates

2. AMPLIFY ON/OFF-TARGET SITE(S)

Cells lysis and PCR with barcoded primers

3. SEQUENCE

Samples pooled and sequenced on Illumina® platform

4. ANALYZE DATA INTERACTIVELY

Visualize results (login required) in genoTYPER-NEXT browser

ADDITIONAL NGS SERVICES FOR CRISPR VALIDATION

STREAMLINE YOUR PATH TO DISCOVERY

Gene-to-Discovery Solutions

AAV Packaging, Custom mRNA Synthesis, Lentivirus Packaging, Recombinant Antibody Production


FEATURES & BENEFITS

Automated, high-throughput workflow – Up to 10,000 samples per run​
Hassle-free - Avoid gDNA extraction, pre-screening prior to sequencing, and TA cloning​
Highly sensitive – Detection of <1% allele frequency, full INDEL resolution, and frameshift analysis
More reasons to choose Azenta

NEW TO OUR PCR PORTFOLIO

Digital PCR (dPCR)

Detect your target sequences with unmatched accuracy

Case Study | Rapid, Scalable, and Ultra-Sensitive Validation of Allele-Specific Gene Editing Events with genoTYPER-NEXT™

Discover the powerful capabilities of genoTYPER-NEXT in this illuminating case study and learn how this high-throughput assay allows rapid and accurate genotyping of CRISPR-edited cell lines.

Webinar | Rapid and Sensitive Validation of Gene Editing with Novel NGS Assays

Learn about current techniques to validate gene edits and how recent advancements in next generation sequencing are improving the sensitivity and scalability of screening genomic manipulations.

Blog | NGS, PCR, or Sanger Sequencing: An Assay Selection Guide

This selection guide offers practical information about PCR + Sanger, qPCR, and NGS approaches to help you determine which assay best suits your project requirements, along with an interactive assay selection tool to aid your decision making.

How To Order


Email | Phone 1-877-436-3949, Ext. 1