ZytoLight^® SPEC CDKN2A/CEN 9 Dual Color Probe

The ZytoLight^® SPEC CDKN2A/CEN 9 Dual Color Probe (PL22) is a direct-labeled FISH probe for the qualitative detection of deletions of the human CDKN2A gene and the detection of the classical satellite III region of chromosome 9 in cytologic or formalin-fixed, paraffin-embedded specimens using fluorescence in situ hybridization (FISH). The probe is intended for use with ZytoLight FISH Implementation Kits.

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 CDKN2A/CEN 9 Dual Color Probe comprises:

• ZyGreen (excitation 503 nm/emission 528 nm) labeled polynucleotides (~10 ng/µl), targeting sequences mapping in 9p21.3* (chr9:21,742,629-22,056,853) harboring the CDKN2A gene region;
• ZyOrange (excitation 547 nm/emission 572 nm) labeled polynucleotides (~1.5 ng/µl), targeting sequences mapping in 9q12 specific for the classical satellite III region D9Z3 of chromosome 9;
• 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 or formalin-fixed, paraffin-embedded specimens to qualitatively detect deletions of the human CDKN2A gene and the detection of the classical satellite III region of chromosome 9, via FISH.

Using suitable filter sets, the hybridization signals of the probe appear green at the CDKN2A, CDKN2B, MTAP gene region and orange at CEN 9.

• Normal scenario: in normal cells during interphase or cells without a deletion involving the gene region CDKN2A, CDKN2B, or MTAP, two green and two orange signals are observed.
• Aberrant scenario: in a cell with deletion affecting the gene region, fewer green signals are observed. Careful observation of the probe localization may be necessary to confirm that partial deletions of the respective gene region will show a normal signal pattern with smaller green signals.

Overlapping signals may appear yellow.

ZytoLight is a registered trademark of ZytoVision GmbH in Germany