PRINS Reaction

1. 2'-Deoxyadenosine 5'-triphosphate (dATP): 100 mMsolution (Roche Diagnostics, Meylan, France) diluted 1:10 with sterile distilled H2O.

2. 2'-Deoxycytosine 5'-triphosphate (dCTP): 100 mM solution (Roche Diagnostics) diluted 1:10 with sterile distilled H2O.

3. 2'-Deoxyguanosine 5'-triphosphate (dGTP): 100 mM solution (Roche Diagnostics) diluted 1:10 with sterile distilled H2O.

4. 2'-deoxythymidine 5'-triphosphate (dTTP): 100 mM solution (Roche Diagnostics) diluted 1:100 with sterile distilled H2O.

5. Fluorescein-12-2'-deoxyuridine 5'-triphosphate (FITC-12-dUTP),1 mM (Roche Diagnostics).

6. Bovine serum albumin (BSA; Sigma, St. Louis, MO).

7. Taq DNA polymerase (Roche Diagnostics) or AmpliTaq (Perkin Elmer, Foster City, CA).

8. 10X Taq buffer: 500 mMKCl, 100 mMTris-HCl, pH 8.3, 15 mMMgCl2 .

9. Oligonucleotide primer specific for chromosome 9, at 50 pmol/|jL (see Table 1 in Chapter 6).

10. Sterile distilled water.

11. Washing buffer: 2X SSC, diluted from 20X SSC in distilled water.

12. 1.5 mL of sterile microcentrifuge tubes (Eppendorf AG, Hamburg, Germany).

13. Cover slips (22 x 32 mm; CML, Nemours, France).

15. Programmable thermal cycler equipped with a flat pate block (Hybaid Ltd., Teddington, UK; see Note 2).

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