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Fiber FISH (CAT#: STEM-MB-1177-WXH)

Introduction

Fiber-FISH is defined as the application of fluorescence in situ hybridization (FISH) techniques on extended DNA molecules. In most cases, the target DNA is prepared by deproteinizing chromosomal chromatin by extraction with detergents or high salt concentration. This treatment removes histones from chromatin and leads to a complete unwinding of the DNA to the level of a linear double helix of approx 340 nm/1000 bp.
Fiber FISH provides a higher resolution of analysis than conventional FISH and yields more precise information as to the localization of a specific DNA probe on the chromosome.
The word "fiber" here refers not to dietary fiber but to a chromosome fiber.




Applications

Determination of numbers of repetitive genes.
Establishing the physical order of cloned DNA fragments along continuous sections of individual chromosomes.
Study DNA replication in yeast artificial chromosomes

Procedure

1. Cells are spread and air-dried on a slide glass.
2. The cells on a slide glass are lysed by the detergent in a closed container.
3. Chromatin fibers attach to the slide glass when it is slowly removed from the container.
4. The preparation is fixed with ethanol.
5. The probes are hybridized following standard procedures.

Materials

• Flow cytometer
• Fluorescence microscopy
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