Detection

1. 1% Avidin-fluorescein DCS (Vector Laboratories, Burlingame, CA) and 1% anti-dig-rhodamine (Roche Molecular Biochemicals; diluted with blocking buffer).

2. 1% Avidin-rhodamine (Roche Molecular Biochemicals) and 1% anti-dig-fluo-rescein (Roche Molecular Biochemicals; diluted with blocking buffer). Either of two detection mixes above can be used according to the selected labelling protocol for multiple-chromosome target detection (see Note 3).

4. Counterstaining solution: In 1 mL of 0.1 MTris-HCl, pH 7.0, dissolve 125 ng of DAPI (Sigma), 1 mg of p-phenylenediamine, and then mix with 9 mL of glycerol.

5. Fluorescence microscope equipped with appropriate filters and connected to an image system is needed (e.g., ISIS 2 in MetaSystems, Belmont, MA).

Was this article helpful?

0 0

Post a comment