dGH Cell Prep Kit

The directional Genomic Hybridization (dGH) Cell Prep Kit is used to prepare actively dividing cells for any of KromaTiD’s dGH assays. 

Product Description

The dGH Cell Prep Kit contains a media additive which incorporates into DNA during replication and allows the daughter strands to be stripped from metaphase chromosomes. This unique method enables high resolution, direct measurement of inversions, translocations, and other structural variations on a cell by cell basis. 

Key Benefits

• Allows processing of your samples in-house
• Fully compatible with your cell culture procedures
• Ready to use with dGH In-Site™ and dGH SCREEN™ assays
• Flexible packaging sizes for processing 10-100 samples
• Orthogonal analysis to PCR/sequencing techniques through direct, single cell visualization of structural variations

dGH Applications

The dGH Cell Prep Kit is used with dGH In-Site™ and dGH SCREEN™ assays for a variety of applications:

• Measuring transgene integration, even from complex or heterogeneous cell populations
• Whole genome structural variant analysis
• Measuring on & off target structural variation in batches of edited cells
• Selecting structurally pure IPSC clones
• Assuring the stability of cell lines

dGH Assays (sold as a separate service)

KromaTiD’s dGH in-Site assays provide whole genome tracking inserting DNA cassettes as small as 2kb. Unidirectional dGH probes provide a single cell, genome-wide perspective of cellular engineering outcomes, including tracking of viral and non-viral mediated insert integration (CRISPR/Cas and alternative systems).

• On- and off-target integration metrics
• Detects structural variations in off target CRISPR therapies
• Clean integration data, even from complex or heterogeneous cell populations
• A unique, whole genome, orthogonal method of direct visualization of inserts, without bioinformatic prediction of outcomes
• Multi-channel fluorescence for flexible and multiplex assay design
• Available for human, murine, canine, non-human primate and CHO cells.

Offering the lowest limit of detection of integrated or genomic DNA targets by fluorescence, dGH in-Site is the most comprehensive tool available for researchers interested in tracking engineering outcomes in a de novo fashion.

dGH SCREEN™ is a single cell assay designed to monitor structural variants throughout the genome in an entirely de novo fashion. By utilizing directional Genomic Hybridization technology combined with fluorescence labeling patterns and chromosomal aggregation strategies, dGH SCREEN™ provides the most comprehensive and high-resolution karyographic analysis available.

Application Benefits of SCREEN™:

• Exchange Events including reciprocal, balanced and allelic translocations;
• Orientation Events including inversions, recombinations and sister chromatid exchanges;
• Chromosomal Gain & Loss Events including sister chromatid fusions, dicentrics/acentrics, fragmentation/chromothrypsis, polypoidy, aneuploidy, monosomy, polysomy;
• Monitoring Cellular Engineering Outcomes: Genome-wide, cell-by-cell and chromosome-by-chromosome assessment of structure, pre- and post modificatio;n
• Orthogonal Data for Sequencing: Genome-wide, confirmatory data regarding rearrangements predicted with long read and other NGS analyses;
• Structural Integrity QC: A straightforward yardstick by which to measure the relative stability of cell lines, or to screen and compare candidate cell lines, based on total genomic structural variation metrics;
• Variant Discovery: Discover previously unknown mutations by de novo assessment of single cells from patient sub-types;
• Genomic Stability Assessment: Track persistence of variants over time, passages, process variable changes, etc.

With dGH SCREEN™, researchers are able to track and localize a wide variety of chromosomal rearrangements within a 10kb limit of detection. In addition to being an unbiased tool for the detection of structural variants throughout the genome SCREEN™ offers High Resolution Analysis of DNA repair activity by monitoring hallmarks of genomic instability, including sister chromatid changes. This analysis produces vast amounts of rearrangement data which can be used to create stability and structural variance profiles for complex and heterogeneous cell populations, making dGH SCREEN one of the most comprehensive and robust karyographic assays available.