How important is the denaturation temperature?
It is vital. The probes which are specific for repeat elements of the human genome are less susceptible to over-denaturation than those specific for single sequences of DNA. They can tolerate much more loss in activity as there are more repeat units to which the probes can hybridise. However, one must also consider the sample. This is made of DNA too, and will lose significant levels of morphology if overdenatured. It is always good practice to take good care when denaturing any FISH product to ensure good signals and good sample morphology. • We are having problems visualising the probes in square 2 (12 centromere) in the CLL panels. We use a Vysis Hybrite heating block. Using Hybrite: Moisten the two strips of paper towel with deionised water (should not be dripping wet) and place the towels in the troughs along the heating surface. Make sure the towels are not protruding above the heating surface and come directly into contact with any slides. Cover spare spaces with empty slides. P
