KromaTiD’s directional Genomic Hybridization™ (dGH) technology combines bioinformatics driven design of unique single-stranded synthetic probes with strand-specific hybridization strategies, and is the only genomics approach capable of detecting DNA sequence, location and orientation in a single test. Assay design with dGH™ provides extremely strong signal strength, eliminates background, and detects small chromosomal rearrangements with high resolution.
For the discovery or detection of inversions, proprietary dGH probes are used with an innovative chromosome preparation in a technique that is capable of determining orientation information from imaging data.
When used as Pinpoint FISH reagents with standard FISH methodologies and equipment, KromaTiD’s dGH probes maximize assay performance by improving kinetics, reducing background and eliminating the need for blocking DNA (COT).
With dGH, researchers are able to directly visualize all manner of genomic structural abnormalities including translocations, sister-chromatid exchanges, and previously undetectable inversions. Check out the library below for demonstrations of real dGH structural rearrangement detection imaging.
Directly reading structure in individual cells resolves disease causing variations that cannot be calculated bioinformatically.
A probe ladder was designed for the antisense strand, and used in combination with Ch 2 chromatid paint to narrow breakpoint region locations. This picture demonstrates a metaphase spread with an inversion.
NFKB2 probe for exons 1-12 on DAPI. Inversion visible in one homolog of Chromosome 10.
Limit of Detection test. Signals corresponding to different sized targets are shown on a single chromosome. Routine detection of a 10Kb target was demonstrated.
Pinpoint FISH probes designed to determine co-location of a binding site to a reference location. The binding site probe (red) is 10KB, and the reference location probe (green) is 100KB.
KromaTiD assays can be run on existing chromosome imaging equipment, and therefore require minimal training and no additional capital investment.
Taking advantage of cutting-edge genomics and bioinformatics strategies, Directional Genomic Hybridization can be efficiently deployed in projects ranging from one to thousands of samples. With the ability to resolve very small inversions and translocations, dGH technology can be employed in both directed and de novo discovery studies, quickly providing data on new or variable rearrangements for your patient screening or target validation needs. If a complete genetic profile is required, pair dGH with NGS on the same samples to build a comprehensive mutation profile.
For inversions or translocations with known breakpoints, targeted dGH assays are an efficient method of screening large populations or libraries, and can be multiplexed and automated to provide data on multiple aberrations in the same test.
Contact us today to talk to our experts about how dGH can solve your research needs!
Replicated Chromosome Metaphase Configuration. Pre-Replication G1 Chromosome. *Inverted Segment. 1) Selective Daughter Strand Degradation 2) Single-Strand dGH Probes 3) Imaging. Visual Orientation Data. When used on metaphase chromosomes, dGH is the only imaging technology capable of providing sequence, location and orientation information in a single assay. The KromaTiD platform enables orders of magnitude higher-resolution inversion detection than any competing technique. Chromosome and Chromatid Assay Format. Using the same probes in different test conditions, KromaTiD assays are capable of targeting entire chromosomes (double-stranded applications like FISH) or individual chromatids (single-stranded applications) in interphase or metaphase cells. Quantitative Mutation Size and Location. Because dGH assays are based on a defined library of sequenced and calibrated genomic probes, they can precisely locate and quantitatively size rearrangements. Unique Specificity. Probes are designed to target unique genomic sequences, so KromaTiD assays require no blocking DNA (COT), exhibit no non-specific background, and demonstrate improved hybridization performance. Single Cell Analysis. dGH assays generate imaging data for single cells, not pools of cells, so are ideal for determining mutation heterogeneity within mixed cell populations while simultaneously identifying recurrent rearrangements. The Broadest Assay Range. In a single assay, KromaTiD products detect the broadest possible spectrum of chromosome rearrangements, including those assayable by standard FISH technologies (e.g. translocations between chromosomes) as well as intra-chromosomal rearrangements such as cryptic inversions.