Chromosome painting

CHROMOSOME PAINTING: A METHOD FOR CELL CHARACTERIZATION

1. CHROMOSOME PAINTING: A METHOD FOR CELL CHARACTERIZATION Presented by: Prince Ph.D. Scholar Enroll. no. 18903011 Department of Biotechnology Indian Institute of Technology, Roorkee

2. INTRODUCTION

3. WHY??????

4. Fluorescent  In-Situ  Hybridization (FISH) identification of Human Chromosomes -   " Chromosome Painting"

5. A technique that hybridizes a DNA nucleic acid probe to a target DNA sequence contained within a cell nucleus. A variety of specimen types can by analyzed using FISH. The intact cells are attached to a microscope slide using standard cytogenetic methods . FISH

6. Probes Probe is a nucleic acid that can be labeled with a marker which allows identification and quantitation . will hybridize to another nucleic acid on the basis of base complementarity .

7. Probes Types of labeling Direct & Indirect Radioactive ( 32 P, 35 S, 14 C, 3 H) Fluorescent Biotin, fluorescein , Digoxigenin etc.

8. FISH for Detection of Single to Multiple Genetic Events Single Target One color Dual Targets Two colors Multiple Targets Multi- colors Allows one to look at multiple genomic changes within a single cell, without destruction of the cellular morphology.

9. PROCEDURE FOR SAMPL E PREPARATION

10. Types of FISH Probes Centromere Telomere Whole chromosome paint locus

11. Types of probes Centromeric (satellite) probe s Locus specific probe s Whole chromosome painting probes

12. Visualization of the Probe DNA probe is labeled with a colored fluorescent molecule. This fluorescent molecule remains attached to the DNA during the hybridization process The molecule emits a particular color when viewed through a fluorescence microscope that is equipped with the appropriate filter sets.

13. Fluorescent Microscope CCD Camera Filters FISH Analysis Software

14. APPLICATION To find out the location of a specific gene located on a specific chromosome. Detection of translocation. Detection of chromosome abnormalities. To find out the chromosomal similarity between divergent species. Prenatal genetic disorder diagnosis.

15. Thanks.