Southern blotting may be undertaken during testing for Fragile X and other triplet repeat expansion disorders.
Southern blotting provides information about whether DNA is absent or present, the size of the DNA and how much DNA is present.
The step-by-step process is outlined below:
- DNA digestion. The DNA is fragmented using a restriction endonuclease enzyme.
- Gel electrophoresis. The DNA fragments are separated by gel electrophoresis. The fragments are separated by size with the smallest moving the furthest through the agarose gel.
- DNA denaturation. The DNA is denatured by immersing the gel in alkali.
- Blotting. The denatured DNA is transferred to a nitrocellulose membrane.
- Probe labelling. The membrane is exposed to a labelled probe that will seek out and hybridise to the complementary DNA fragment. After hybridisation, excess probe is washed from the membrane.
- Detection. If a radiolabelled probe is used, visualisation is by X-ray, or if a chromogenic detection method is used it is visualised by the development of colour on the membrane.
Source:-https://www.futurelearn.com/courses/molecular-techniques/6/steps/765691
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