Introduction

Northern blot, also called RNA blot, is a technique used to detect the specific RNA molecules among a mixture of RNA. In molecular biology, it could be used to analyze a specific RNA from a particular sample (tissue or cell) in order to measure the RNA expression of particular genes.

In Northern Blot, the RNA samples weas first separated by size via electrophoresis in a gel under denaturing conditions, then the samples were transferred to a membrane. On the membrane, the RNA were cross-linked and hybridized by a labeled probe which contains a sequence that is complementary to a particular RNA sequence in the sample. After hybridization, the RNA with the probe were detected from other RNA molecules on the membrane.

Procedures:

1. Isolation total RNA or mRNA from tissue samples.

2. Fractionation of RNA through agarose gel.

3. Transfer RNA onto a supercharged membrane.

4. Cross-liked RNA by UV light.

5. Hybridized membrane with labeled probes.

6. Visualization of the labeled RNA on X-ray film.

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