Identification of microorganisms by Fluorescence in situ hybridisation (FISH) (CAT#: STEM-MB-1185-WXH)

Introduction

Since their first application as ‘phylogenetic stains’ in 1989, fluorescently labelled, rRNA-targeted oligonucleotide probes have become a common tool for the direct, cultivation-independent identification of individual bacterial cells. Fluorescence in situ hybridisation (FISH) with rRNA-targeted oligonucleotide probes has been developed for the in situ identification of individual microbial cells and is now a well-established technique.

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Principle Applications Procedure Materials Notes Related Products

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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