ZytoLight^® SPEC TERT/5q31 Dual Color Probe

The ZytoLight^® SPEC TERT/5q31 Dual Color Probe (PL50) is a direct-labeled FISH probe intended for the qualitative detection of amplifications involving the human TERT gene and for detecting specific sequences on chromosome 5q31 in formalin-fixed, paraffin-embedded (FFPE) specimens. It is designed for use with the ZytoLight FISH-Tissue Implementation Kit.

Principle:

The fluorescence in situ hybridization (FISH) technique facilitates the detection and visualization of specific nucleic acid sequences. Fluorescently-labeled DNA fragments (FISH probes) co-denature and hybridize with complementary target DNA strands. Following stringent washing to remove unbound fragments, the hybridized probes are visualized with a fluorescence microscope that utilizes specific excitation and emission filters.Fluorescence in situ hybridization (FISH) uses direct-labeled probes that co-denature (75°C/10 min), hybridize overnight (37°C) to FFPE targets, and are stringency-washed for visualization of green (TERT)/orange (5q31) signals via specific filters after DAPI counterstaining. ZytoLight^® eliminates antibody detection for aberrations like amplifications

Components:

ZytoLight® SPEC TERT/5q31 Dual Color Probe comprises:

• ZyGreen labeled polynucleotides (excitation 503 nm/emission 528 nm), targeting sequences in the TERT gene region on chromosome 5 (5p15.33).
• ZyOrange labeled polynucleotides (excitation 547 nm/emission 572 nm), targeting specific sequences in the 5q31 locus.
• Formamide-based hybridization buffer.

For the qualitative detection of human TERT gene amplifications and chromosome 5q31 specific sequences by fluorescence in situ hybridization (FISH)

The probe is designed for qualitative detection of sequences in the TERT gene region and chromosome 5q31 specific sequences in formalin-fixed, paraffin-embedded (FFPE) tissue specimens by FISH. It is used in combination with the ZytoLight^® FISH-Tissue Implementation Kit for specimen pretreatment, hybridization, and post-hybridization processing.

In a normal interphase nucleus, two green signals (for TERT) and two orange signals (for 5q31) are expected. In cells with TERT amplifications, an increased number of green signals or clusters may be observed. Any unexpected signal patterns should be further investigated to identify potential genetic rearrangements.

ZytoLight is a registered trademark of ZytoVision GmbH in Germany