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If only one chromatid owns a detectable spot (Fig 4E), write a zero value and indicate ss for single spot at the corresponding location in « Table 1 ». Choose another chromosome and increase the chromosome number by 1. Write a line from the previous chromosome to the new one (this will be helpfull at the karyotyping step) on the printed metaphase. Perform the spot measurement for the new chromosome as in step 8 (Fig 4D ,E) Repeat the operation until the last chromosome is processed. Save the results of the measurements with the following file name « Slide-MetaphaseN results-NIH », where « Slide » and « MetaphaseN » are the names of the slide and the number of the metaphase analyzed respectively.

BSA (fraction V; Serva) teleostean gelatin (Sigma). 5. Biotinylated goat anti-avidin antiserum (Vector Labs, Burlingame, CA). 6. Water bath, preferably with shaking. Used to adjust the temperature of the solutions in Coplin jars. Always measure temperatures inside Coplin jars. 9. Reagents for Specimen Counter Staining 1. Propidium iodide (PI; Sigma). Dissolve at 1 mg/mL in sterile water, store at –20°C. 2. DAPI (4',6-diamidine-2'-phenylindole dihydrochloride) (Sigma), 1 mg/mL in sterile water; store at –20°C.

Copy the intensity value, corresponding to ds elements, from column 5 to column 10. 12. Divide the values contained in column 3 by those in column 9 and store the result in column 15. 13. Divide the values contained in column 5 by those in column 10 and store the result in column 16. 14. Select columns 9 and 10, the columns containing the sum of the intensity of the spots, and calculate the statistical parameters mean and standard deviation (SD), with the command Calc/Statistics; select the Selected cells option and (at least) the Basic information option.

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