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Location: Home > Custom Services > Immunology Services > In Situ Hybridization

In Situ Hybridization

Date: 2018-01-25 Author: Leading Biology Click: 2485

Introduction

In situ hybridization (ISH) is a type of hybridization that uses a labeled complementary DNA, RNA or modified nucleic acid strand which allows for precise localization of a specific segment of nucleic acid within a histologic section. The underlying basis of ISH is that nucleic acids, if preserved adequately within a histologic specimen, can be detected through the application of a complementary strand of nucleic acid to which a reporter molecule is attached. Visualization of the reporter molecule allows to localize DNA or RNA sequences in heterogeneous cell populations including tissue samples and environmental samples, Riboprobes also allow to localize and assess the degree of gene expression. 


There are four types of ISH probes: Double-stranded DNA (dsDNA) probes, Single-stranded DNA (ssDNA) probes, RNA probes (riboprobes), and Synthetic oligonucleotides (PNA, LNA). The probes can be labeled both by radioactive isotopes (e.g. 32P, 35S, 3H) and non-radioactive labels (e.g. biotin, digoxigenin, fluorescent dye (FISH)).


In Situ Hybridization


FISH (Fluorescence In Situ Hybridization) is a method based on the in situ hybridization techniques and consists in injecting reacts and fluorescents probes into samples to determine location and expression of specifics genes.

Procedures:
1. Sample preparation
2. Co-denaturation and hybridization
3. Wash off of unbound probe

4. Detection and data analyze



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