What temperature is necessary for simultaneous denaturation in FISH procedures?

In Fluorescence In Situ Hybridization (FISH) procedures, simultaneous denaturation of nucleic acid strands is essential to ensure that the probe can hybridize effectively to the target sequences. The typical temperature for this denaturation process is around 90 degrees Celsius. At this temperature, the hydrogen bonds that hold the DNA strands together break down, allowing the strands to separate. This denaturation step is crucial as it creates single-stranded DNA, which is necessary for the probes to bind specifically to the complementary target sequences.

Temperatures lower than 90 degrees Celsius may not be sufficient to fully denature the DNA, potentially leading to incomplete hybridization or non-specific binding in subsequent steps of the FISH procedure. Hence, using 90 degrees Celsius is optimal for achieving reliable and accurate results in FISH applications.