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Determination of Imbalances of chromosome 17 in medulloblastomas by Fluorescence in situ hybridisation (FISH) (CAT#: STEM-MB-1221-WXH)

Introduction

Medulloblastomas make up more than 90% of cases of central nervous system (CNS) primitive neuroectodermal tumours, which are the most common CNS tumours in childhood.
The analysis of genetic abnormalities has been one focus of research on the biology of primitive neuroectodermal tumours. Cytogenetic studies of medulloblastoma have identified several non-random chromosomal aberrations in a high proportion of cases. Of these, isochromosome 17q (i(17q)) is the most common, occurring in a third of successfully karyotyped cases. Microsatellite analysis has corroborated the existence of chromosome 17 abnormalities by documenting the loss of heterozygosity (LOH) on 17p.




Principle

FISH uses fluorescent probes with complementary base sequences to locate the presence or absence of specific portions of DNA on chromosomes. The probe and target DNA must be denatured with heat or chemicals to break hydrogen bonds in the DNA and to allow hybridisation to occur once the two samples are mixed. The fluorescent probes form new hydrogen bonds with their complementary base pairs on the DNA, and these can then be detected via microscopy.

Applications

Detect and localize the presence or absence of specific DNA sequences on chromosomes.
Detect and localize specific RNA targets (mRNA, lncRNA and miRNA) in cells, circulating tumor cells, and tissue samples.

Procedure

1. Sample preparation
2. Co-denaturation and hybridization
3. Probe detection
4. Wash off of unbound probe
5. Analysis by flow cytometer/fluorescence microscopy

Materials

• Flow cytometer
• Fluorescence microscopy
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