Technique / Histology / In situ hybridization

In situ Hybridization techniques


In situ Hybridization techniques

In situ hybridization (ISH) can be used to detect nucleic acid sequences in tissue and cell cultures. Its applications include detection of mRNA expression in situ, chromosomal rearrangement, viral RNA expression in host cell etc.

General steps for in situ hybridization:

1. Probe synthesis by gene cloning, PCR or oligo synthesis and labeling with radioisotope or DIG.

2. Preparation of fixed tissue (embedding and sectioning) or chromosome spreads.

3. Pretreatment with proteinase K to increase probe accessibility.

4. Hybridization of probe and tissue target sequences.

5. Washing and detection.

in situ hybridization introduction and protocol (from GeneDetect)

Issues discussed in the technical note:

Introduction
Preparation of material
Choice of probes
Probe types
Benefits of using oligonucleotide probes
Labeling your oligonucleotide
Hybridization Issues
Controls
Detailed Protocols for Download

Whole-mount in situ hybridization illustration

Last update 07-Feb-2008, Rating Very Good of 2 votes.

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By Dr. R.K.Chahota on 28-Dec-2008
The information is very good from introduction part to advanced version of FISH technique.
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By Edder Mazariegos on 07-Feb-2008
Tengo problemas para la hibridación in situ, me dan resultados positivos y luego negativos en los controles. Que hago?? Que tan estable es el reactivo despues de la fecha de caducidad???
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