ZytoLight^® SPEC D13S319/13q34 Dual Color Probe

The ZytoLight^® SPEC D13S319/13q34 Dual Color Probe is a direct-labeled FISH probe designed for the qualitative detection of deletions in the human D13S319 region and chromosome 13q34 sequences in cytology specimens through fluorescence in situ hybridization (FISH). The probe is intended for use with the ZytoLight FISH-Cytology 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 D13S319/13q34 Dual Color Probe comprises:

• ZyOrange (excitation 547 nm/emission 572 nm) labeled polynucleotides (~4.5 ng/µl), targeting sequences mapping in 13q14.2* (chr13:50,607,438-50,781,256) harboring the D13S319 locus;
• ZyGreen (excitation 503 nm/emission 528 nm) labeled polynucleotides (~10.0 ng/µl), targeting sequences mapping in 13q34* (chr13:113,691,216-114,323,467);
• 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 cytologic specimens to qualitatively detect deletions involving the human D13S319 region and chromosome 13q34 specific sequences, via FISH.

Using suitable filter sets, the hybridization signals of the probe appear orange at the D13S319 locus and green at the 13q34 locus.

• Normal scenario: in interphases of normal cells or cells without a deletion of the D13S319 locus, two orange signals and two green signals are observed.
• Aberrant scenario: in cells with deletion affecting the D13S319 locus, fewer orange signals will be observed. Partial deletions of the D13S319 locus might result in a normal signal pattern with smaller orange signals. In cells with deletions affecting the 13q34 locus, fewer green signals will be observed.

Overlapping signals may appear yellow.

ZytoLight is a registered trademark of ZytoVision GmbH in Germany