ZytoLight^® SPEC USP6 Dual Color Break Apart Probe

The ZytoLight^® SPEC USP6 Dual Color Break Apart Probe is a direct-labeled fluorescence in situ hybridization (FISH) probe designed for the qualitative detection of translocations involving the USP6 gene at 17p13.2. Optimized for use with formalin-fixed, paraffin-embedded (FFPE) tissue samples, this probe enables precise identification of genetic rearrangements through fluorescence in situ hybridization (FISH). For best results, the probe should be used in conjunction with the ZytoLight^® FISH-Tissue Implementation Kit.

ZytoLight^® probes enable the precise identification of genetic abnormalities, including translocations, deletions, amplifications, and chromosomal aneuploidies, in formalin-fixed, paraffin-embedded (FFPE) tissue sections or cytology specimens, without the need for additional antibody-based detection.

Components

The ZytoLight SPEC USP6 Dual Color Break Apart Probe comprises:

• ZyGreen (excitation 503 nm/emission 528 nm) labeled polynucleotides (~10.0 ng/µl), targeting sequences mapping in 17p13.2* (chr17:4,489,889-5,017,582) distal to the USP6 breakpoint region;
• ZyOrange (excitation 547 nm/emission 572 nm) labeled polynucleotides (~4.5 ng/µl), targeting sequences mapping in 17p13.2* (chr17:5,087,046-5,361,104) proximal to the USP6 breakpoint region;
• Formamide based hybridization buffer

*according to Human Genome Assembly GRCh37/hg19

Principle

The fluorescence in situ hybridization (FISH) method enables the identification and visualization of particular nucleic acid sequences within cell samples. DNA fragments labeled with fluorescent dyes, known as FISH probes, are co-denatured with their complementary target DNA strands in the sample and then allowed to hybridize. Following hybridization, non-specific and unbound probe fragments are removed through stringency washes. The DNA is then counterstained with DAPI, and the hybridized probes are detected using a fluorescence microscope equipped with filters matched to the specific fluorochromes used to label the FISH probes.

This probe is intended for use in formalin-fixed, paraffin-embedded specimens to qualitatively detect translocations involving the USP6 gene at 17p13.2, via FISH.

Using suitable filter sets, the probe’s hybridization signals are observed as green for regions distal to the USP6 breakpoint and orange for regions proximal to it.

• Normal scenario: in normal cells or those without a USP6 gene translocation, two green/orange fusion signals are visible during interphase.
• Aberrant scenario: if a translocation affects the USP6 gene region, this is shown by one distinct green signal and one distinct orange signal.

Overlapping orange and green signals may appear yellow.

ZytoLight is a registered trademark of ZytoVision GmbH in Germany