Researchers provide edited cell line(s) of interest.
CRISPR edited cell lines submitted to GENEWIZ in 96-well plate form.
Direct cell lysis, and 2-step PCR amplification with barcoded primers.
Barcoded samples pooled for sequencing on Illumina platform.
Wells containing correctly targeted cells are visualized in the genoTYPER-NEXT data browser..
(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.
(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.
(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.
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.