Southern Blotting Flashcards
What does Southern Blotting do?
It combines gel electrophoresis and nucleus acid hybridization.
What does Southern Blotting allow us to do?
It allows us to detect just those bands that include parts of the B-globin gene.
What is the first step?
Mixing each DNA sample with the same restriction enzyme.
What does the digestion of each sample yield?
A mixture of thousands of restriction fragments.
What does electrophoresis do?
It separates the restriction enzymes.
What does the separation of restriction fragments form?
A characteristic pattern of bands.
What does capillary action do?
It pulls the alkaline solution upward through the gel.
What does the the pulling of the alkaline solution upward through the gel cause?
The transfer of DNA to a nitrocellulose membrane. (Producing the blot)
What happens during the process of transferring DNA to a nitrocellulose membrane?
DNA is denatured.
What is positioned in bands corresponding to those on the gel?
The single stands of DNA stick to the nitrocellulose.
Next, nitrocellulose is exposed to what?
A solution contains a radioactively labeled probe.
What is unique about the probe?
It is single-stranded DNA complementary to the B-globin gene.
How can probe molecules attach?
By base pairing to any restriction fragments containing a part of the B-globin gene.
What is placed over the blot?
Photographic film.
What happens in the bound probe?
The radioactivity exposes the film to form an image corresponding to those bonds that contain DNA.
