genoTYPER-NEXT

Recent advancements in genome editing techniques have allowed researchers to choose from multiple molecular approaches to achieve targeted DNA modifications in living organisms and cells. Current techniques leverage new and historical systems, such as CRISPR/Cas9 nucleases, transcription activator-like effector-based nucleases (TALENs), and zinc finger nucleases (ZFNs) to achieve increasingly precise gene editing results. Despite the ability to perform high-throughput targeting, identification of correctly targeted cells remains a significant bottle neck for large-scale projects.

GENEWIZ has developed genoTYPER-NEXT, a proprietary NGS-based application ideal for ultra-sensitive, high-throughput, sample-to-answer genotyping projects. This tool provides an effective and efficient option to screen your targeted cell lines through an automated, interactive, and intuitive platform


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FEATURES & BENEFITS


Complete sample-to-answer solution starting from cells



Automated, high-throughput workflow - Up to 10,000 samples per run


Hassle-free - Avoid gDNA extraction, pre-screening prior to sequencing, TA cloning, etc


Sensitive - <1% minor allele frequency detection, full INDEL resolution, detection of unique molecules, and frameshift analysis

SAMPLE TO ANSWER

SAMPLE

1. GENOME EDITING

Researchers provide edited cell line(s) of interest.

SAMPLE

2. CLONES IN 96 WELL PLATES

CRISPR edited cell lines submitted to GENEWIZ in 96-well plate form.

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3. PCR PRIMER DESIGN

Direct cell lysis, and 2-step PCR amplification with barcoded primers.

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4. SEQUENCING

Barcoded samples pooled for sequencing on Illumina platform.

ANSWER

5. GENOTYPER NEXT DATA BROWSER

Wells containing correctly targeted cells are visualized in the genoTYPER-NEXT data browser..

Comparison

Deliverables

Figure 1. Reads distribution on targeted region.


(A) Scatter plot showing sequencing depth across 10 x 96 plates. (B) Visualization of a representative well demonstrating ~90% of reads cover desired target region. (C) Overview of mutation frequency across 10 x 96 plates.

Figure 2. Tunable and interactive filtering of clones.


(A) User-defined thresholds for homozygous mutant and wild type may be entered in the provided HTML file and are automatically applied to the selected plates(s). (B) By tuning the homozygous mutant threshold parameter wells containing correctly targeted of carious purities may be easily identified.

Figure 3. Interactive data analysis module.

(A) Identity of clones illustrated by color-coded wells. For each well, percentage of reads carrying mutations is indicated. When a well is clicked, the module displays sites and frequency of mutations on the amplicon (B) and target region (C). (D) Type and size of INDELs when compared to provided reference sequence. (E) Shows SNP analysis in the target region. DNA (F) and amino acid (G) alignments to reference for the target region.

Sample Submission Guidelines

GENEWIZ accepts frozen cell pellets (preferred) and purified genomic DNA as starting material for genoTYPER-NEXT projects. We can also accept other custom starting materials for extraction upon request. For detailed sample submission requirements please visit our Sample Submission Guidelines


How To Order


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