Unlock Exclusive Discounts & Flash Sales! Click Here to Join the Deals on Every Wednesday!

Analysis of Trypanosoma noyes by Fluorescence in situ hybridisation (FISH) (CAT#: STEM-MB-1213-WXH)

Introduction

Trypanosomes are global protozoan parasites that infect all classes of vertebrates and are typically transmitted by haematophagous invertebrate vectors. In Australia, an increasing number of novel Trypanosoma species have been identified from various wildlife hosts, some of which are critically endangered. Trypanosoma noyesi is a recently described species of biosecurity concern, due to a close relationship to the South American human pathogen, Trypanosoma cruzi. This genetic similarity increases the risk for introduction of T. cruzi via a local vector.




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
Advertisement