American Society for Clinical Pathology (ASCP) Technologist in Cytogenetics certification (ASCP CG) Practice Test

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What temperature is necessary for simultaneous denaturation in FISH procedures?

80oC
90oC
100oC
95oC

The correct answer is: 90oC

The necessary temperature for simultaneous denaturation in FISH (Fluorescence In Situ Hybridization) procedures is 80°C. This temperature is critical because it is sufficient to break the hydrogen bonds between the complementary DNA strands, allowing them to separate and become single-stranded. This enables the fluorescent probes to hybridize effectively with their target sequences on the chromosomes. While other temperatures, such as 90°C, 100°C, and 95°C, might also lead to denaturation, they exceed the optimal condition necessary for FISH. Temperatures that are too high can lead to excessive denaturation, potentially affecting the stability of the probes and their binding accuracy. Therefore, sticking to around 80°C is ideal for obtaining clear and reliable results in FISH experiments.